molecular biology and genetics …however, urgent laboratory investigations are undertaken when...

MOLECULAR BIOLOGY AND GENETICS

LABORATORY

USER GUIDE

This manual is designed to give an overall view of the services available in MBG Lab. It is intended

as a quick reference guide. This manual will be updated periodically, therefore any unauthorised

printed copies are uncontrolled and care should be taken to ensure that the current version is

utilised.

MBG Lab User Manual

Document ID: MBG-MA3 Issue Number: 1 Revision Number: B Effective Date: 08 Jun 2016 Authorized By: Kamal Khazanehdari

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This is an official document of MBG Lab; the saved or printed copy of this document shall be treated as “uncontrolled”. Always refer to the controlled version.

MBG Lab P.O. Box 597, Dubai, UAE T: +971 4 3372529 F: +971 4 3372769 E: [email protected] Web: www.mbg.ae


Table of Contents

1 LABORATORY INFORMATION .......................................................................................................4

1.1 INTRODUCTION ................................................................................................................................. 4

1.2 LOCATION .......................................................................................................................................... 4

1.3 CONTACT INFORMATION .................................................................................................................. 4

1.4 WORKING HOURS ............................................................................................................................. 4

1.5 ROUTINE REQUESTS .......................................................................................................................... 4

1.6 URGENT REQUESTS ........................................................................................................................... 4

1.7 EMERGENCY/ CRITICAL REQUESTS ................................................................................................... 5

1.8 OUT OF HOURS REQUESTS ................................................................................................................ 5

1.9 ACCREDITION/ LICENSURE ................................................................................................................ 5

1.10 CONFIDENTIALITY.............................................................................................................................. 5

2 SCOPE OF SERVICES PROVIDED .....................................................................................................6

3 SENDING A SAMPLE .....................................................................................................................6

3.1 COMPLETING A REQUEST FORM ...................................................................................................... 6

3.2 SERVICE AGREEMENT ........................................................................................................................ 7

3.3 SAMPLE COLLECTION ........................................................................................................................ 7

3.4 SAMPLE TRANSPORT ......................................................................................................................... 7

3.5 INFECTION HAZARDS AND HIGH-RISK SAMPLES ............................................................................... 8

3.6 ACCEPTANCE AND REJECTION CRITERIA ........................................................................................... 8

3.7 TURN-AROUND TIME ........................................................................................................................ 9

3.8 FACTORS THAT AFFECT THE PERFORMANCE OF THE ASSAY ............................................................ 9

3.9 REQUESTING ADDITIONAL EXAMINATIONS .................................................................................... 10

4 RESULTS .................................................................................................................................... 10

5 REPORTING EMERGENCY/CRITICAL OR URGENT RESULTS ............................................................ 10

6 FEEDBACK .................................................................................................................................. 11

7 COMPLAINTS ............................................................................................................................. 11

8 CUSTOMER SATISFACTION ......................................................................................................... 11

9 VETERINARY MOLECULAR DIAGNOSTIC SERVICES ....................................................................... 11

9.1 Pathogen Identification Assays ....................................................................................................... 11

9.2 Avian Sexing .................................................................................................................................... 11

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9.3 Animal Genotyping ......................................................................................................................... 11

9.4 Paternity Verification ...................................................................................................................... 12

9.5 Species identification ...................................................................................................................... 12

9.6 Avian Species Differentiation .......................................................................................................... 12

9.7 Equine Coat Colour ......................................................................................................................... 12

10 MEDICAL MOLECULAR DIAGNOSTIC SERVICES ............................................................................ 12

Pathogen Identification Assays ................................................................................................................... 12

11 CYTOGENETICS ........................................................................................................................... 12

11.1 Classical Karyotyping ....................................................................................................................... 13

11.2 Fluorescent in Situ Hybridization .................................................................................................... 13

11.3 Sperm Viability Test ........................................................................................................................ 13

12 REGENERATIVE STEM CELL THERAPY .......................................................................................... 13

12.1 MBG-Stem ....................................................................................................................................... 13

12.2 Platelet – Rich Plasma ..................................................................................................................... 14

12.3 Banking ............................................................................................................................................ 15

13 MOLECULAR GENOMIC TECHNIQUES .......................................................................................... 15

18.1 STR Analysis ..................................................................................................................................... 15

18.2 Sequencing ...................................................................................................................................... 15

18.3 SNP Genotyping .............................................................................................................................. 16

18.4 Gene Expression Profiling ............................................................................................................... 16

18.5 Construction of genomic and cDNA libraries .................................................................................. 16

14 RESEARCH AND DEVELOPMENT .................................................................................................. 16

15 EDUCATION AND TRAINING ....................................................................................................... 17

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1 LABORATORY INFORMATION

1.1 INTRODUCTION

The Molecular Biology and Genetics Laboratory (MBG Lab) is a modern facility designed to support an

extensive and diverse range of scientific research. It includes large testing areas with state-of-the art

equipment, safety features and office space. Our interests mainly surround the field of molecular biology,

genetics, cell biology and genomics.

1.2 LOCATION

Address: Molecular Biology and Genetics Laboratory

17th Street, 2ndZa’abeel Road

Dubai, UAE

P.O Box: 597, Dubai, UAE

1.3 CONTACT INFORMATION

Advice or information on the list of assaysoffered by MBG Lab can be provided by contacting the

laboratory in a number of ways:

Tel: +9714 337 2529

Fax: +9714 337 2769

Email: [email protected]

Information on services provided and contact numbers is also available on the Lab’s web site:

www.mgb.ae

1.4 WORKING HOURS

The laboratory is operational on Sundays – Thursdays, 7:00 am – 4:00 pm.

Please refrain from sending samples outside the working hours unless in case of emergency.

1.5 ROUTINE REQUESTS

Routine samples which arrive in the laboratory after 11:00 am will not be processed until the next working

day. Turn-around times for routine assays can be found in Section 10.6

1.6 URGENT REQUESTS

Please request assays to be performed urgently only when it is clinically essential. All of our work is

processed rapidly and the results are available in a timely manner. Turn-around times for each assay are

published in this user guide in section 10.6

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If you wish for a sample to be analysed urgently, please contact the lab beforehand so that the necessary

arrangements can be made. Also, make sure that the request form clearly states urgent. These samples

will be handled separately and the result reported to the official contact person as soon as possible. (Note:

urgent requests may incur additional charges).

1.7 EMERGENCY/ CRITICAL REQUESTS

Critical/ Emergency requests demand rapid processing and prioritization will be used to process, analyse

and report results. In case of emergency, please contact the Lab as far in advance as possible so that we

can set up an emergency protocol suitable for your requirements.

1.8 OUT OF HOURS REQUESTS

At present, no services are routinely offered outside of normal working hours. However, urgent laboratory

investigations are undertaken when considered to be critical and are described in Section 1.7.

1.9 ACCREDITION/ LICENSURE

The Molecular Biology and Genetics Laboratory (MBG Lab) has been accredited by Dubai Accreditation

Centre (DAC) under the recognition of the International Laboratory Accreditation Cooperation (ILAC) to

international standards; ISO15189:2012 and ISO17025:2005. This demonstrates our continuing

commitment to achieve the highest quality service for our customers.

MBG Lab services undergo continuous review through quality assurance and audit activities. The Lab is

committed to performing its activities in accordance with the requirements including:

• Staff recruitment, training, development and retention at all levels to provide a full and effective

service to its users.

• The proper procurement, validation and maintenance of equipment and other resources as are

needed for the provision of the service.

• The collection, transport and handling of all samples in such a way as to ensure the correct

performance of laboratory examinations.

• The use of accredited and validated examination procedures and assay methods that will ensure

the highest achievable quality of all assays performed.

• The reporting of results in ways which are timely, accurate and clinically useful.

• The identification, documentation, investigation and resolution of anomalies.

• The assessment of user satisfaction surveys, in addition to internal audit and external quality

assessment, in order to produce continual quality improvement.

1.10 CONFIDENTIALITY

Confidentiality of Lab results and information are maintained at all times. Sensitive client information is

protected from unauthorized access at all times through the use of password protected secure LIMS

system.MBG Lab is committed to maintaining the confidentiality of all patient information. MBG Lab has

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established a Confidentiality and non disclosure policy to protect the confidentiality of information about

clients. All staff are obliged to sign a legally binding confidentiality agreement upon commencement of

their employment at MBG lab.

2 SCOPE OF SERVICES PROVIDED

Molecular Diagnostics

Molecular Genomics

Next Generation Sequencing

Molecular Cytogenetics

MBG- Stem - Adult Stem Cell Technology

Research and Development

Education And training

Kindlyrefer to sections 9 to 13 for more details of the services provided by the MBG Lab and refer to the

attached Veterinary Service List/ Medical Service Listfor complete list of assays provided. Alternatively,

please refer to our website (www.mbg.ae) for more information.

3 SENDING A SAMPLE

MBG Lab does not offer and clinical advice on ordering of examinations or the interpretation of the results.

Kindly consult your Veterinarian / Clinician to find out which assay you require to be tested for.

3.1 COMPLETING A REQUEST FORM

Request forms are available by contacting the lab using the information in Section 1.3 (Contact

Information). They are also available on the lab website www.mgb.ae.

An appropriate request form must accompany each sample and it is essential that it is filled out legibly and

completely before samples can be processed. The request form must have the following

essentialinformation:-

Unique identity of the patient including gender, date of birth, and location/Contact Details

Name/unique identity of the authorized requester, along with their address

Means and destination for reporting the results (Name, maid id, Fax No. etc)

Type of sample (and anatomic site of origin where appropriate)

Examinations requested

Clinical information relevant to the patient

Identity of the person collecting the primary sample & Date and time of primary sample collection

Date and time of receipt of samples by laboratory*

* Will be filled in by the Lab

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3.2 SERVICE AGREEMENT

The contractual arrangement between the laboratory and its users is defined by the laboratory requisition

form. Each request form (together with its relevant primary samples) is checked for conformity with the

laboratory’s test requirements which are made available to users via thewww.mbg.ae. The requisition

form must be filled out legibly and completely to avoid any delay with the testing. By completing a

requisition form the user is agreeing to the terms and conditions provided in this user manual.

3.3 SAMPLE COLLECTION

The best results are obtained when an appropriate, well-taken sample, in the proper container, is delivered

to the laboratory promptly and relevant clinical information provided on the request form.

Kindly refer to MBG Lab website for more details on the appropriate sample types and containers for the

assay required. Please contact the lab if there is any doubt about the best sample to take or you have

questions about any assay.

Great care must be taken in obtaining samples. Equipment such as needles and blades must be

immediately disposed of safely in locally approved "sharps" bins and NOT SENT TO LABORATORY.

Instruction for collection activities

Determine and verify identity of the patient.

Verify that the patient meets any pre examination requirements. (Sample Collection, storage,

transport Guideline-Veterinary/ Sample Collection, storage, transport Guideline-Medical)

Ensure the sample type to be collected is appropriate for the assay.(Veterinary Service List/

Medical Service List)

Samples for certain assays must be collected and transported in sample tubes containing transport

medium. Kindly refer to (Veterinary Service List/ Medical Service List) to know which assays have

special requirements.

Use collection containers appropriate to the sample type being collected (Sample Collection,

storage, transport Guideline-Veterinary / Sample Collection, storage, transport Guideline-

Medical)

Label the sample containers with at least 2 of the unique identifies (Name, unique ID number, Date

of birth) for their appropriate identification and quick processing by the lab

Identity of the person collecting the sample must be recorded along with collection date and time.

Refer to Sample Collection, storage, transport Guideline-Veterinary / Sample Collection, storage,

transport Guideline-Medicalfor proper storage conditions of samples after collection.

3.4 SAMPLE TRANSPORT

To ensure appropriate processing of samples, the requester must ensure that:

Samples are transported at the temperature specified in Sample Collection, storage, transport

Guideline-Veterinary/ Sample Collection, storage, transport Guideline-Medicalto ensureintegrity

of the sample.

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Samples are transported to the laboratory without delay and within the time frame specified.

Samples are transported in a closed container to ensure safety of the carrier, public and receiving

laboratory.

3.5 INFECTION HAZARDS AND HIGH-RISK SAMPLES

It is the responsibility of those sending samples to the lab to convey to the lab suitable and sufficient

information of any potential hazards. Please help to minimize the risk to lab staff by:

Discarding cracked tubes or those with tops off,

Avoid overfilling and contaminating the outside of containers, and

Make sure that tubes and bottles are securely sealed.

It is important that samples from patients known or suspected to be infected with dangerous pathogens

such as blood borne viruses (BBV) are appropriately labelled. The use of “Danger of Infection” or

“Biohazard” label (as shown below) would be appropriate with sufficient detail on the request form to

identify the nature of the risk. The sample and request form may be placed in an envelope to maintain

patient confidentiality whilst in transit. Failure to properly label a sample may result in the analysis being

delayed or refused.

3.6 ACCEPTANCE AND REJECTION CRITERIA

Acceptance criteria:

Samples shall be accepted only if:

The request form contains all the required information, filled legibly

Sample is clearly labelled with at least 2 unique identifiers

The sample type and volume are as recommended

The sample is collected in the recommended container type

The sample is stored and transported as recommended

The sample arrives at lab within recommended time frame after collection.

Rejection Criteria:

The receipt of unsuitable samples may result in either sample rejection or delays in testing/ results. The

sample(s) may be rejected if there is:

• No label, illegible label or it is missing unique identifiers.

• Missing request form.

• Discrepancy between information on the container and/or request form.

• The wrong type of sample container and/or it is broken or damaged.

• Incorrect/insufficient sample received or sample deemed too old for analysis.

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A rejection Form shall be filled for the rejected samples, and is send to the client along with a copy of

request form, stating the reason for rejection.

Exclusions to acceptance/rejection criteria

Exclusions to acceptance and rejection criteria exist and include the following:

The primary sample is irretrievable or critical

Repeat sampling may cause the patient undue pain, stress or inconvenience.

Extra consideration is also given to samples from bleeding patients, paediatrics, geriatrics and

palliative care patients.

In such cases, the report shall clearly state the deviation from the recommendation.

3.7 TURN-AROUND TIME

Turn-around Time offers the user a time-frame (from the receipt of the samples by the lab) within which

the Report of Results will be generated. A general outline of the turn-around times for the various services

offered by MBG Lab is provided in Table 2.

Table2. List of Different Services and their Turn-Around Times for Normal Samples as well as Urgent or

Emergency/Critical Samples

Assay Type

Normal Turn-Around Time

(working days)

Urgent or Emergency/Critical

(working days)**

Pathogen Identification Not exceeding 5 working days Not exceeding 3 working days

Multiplex-PCR based tests Not exceeding 5 working days Not exceeding 3 working days

Avian Sexing & Avian Species

Identification

Not exceeding 5 working days Not exceeding 3 working days

Animal Genotyping & Paternity

Verification

Not exceeding 10 working days -

Cytogenetics Contact MBG Lab Contact MBG Lab

Regenerative Stem Cell Therapy Not exceeding 2 working days Not exceeding 1 working days (if sample arrives before 9:00am)

**Incase of Urgent Samples, samples received before 11am only will be processed on the same day.

Samples received after 11am will be processed the next working day.

Kindly refer to the attached Veterinary Service List/ Medical Service Listfor list of assays provided or refer

to our website.

3.8 FACTORS THAT AFFECT THE PERFORMANCE OF THE ASSAY

There are a number of factors that can affect the performance of the assay resulting in spurious results. These are as follows:

Sample type provided is not recommended

Sample collection For RNA testing is not done in RNA transport media

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Samples are not stored or transported under recommended temperature.

Samples are not delivered within the recommended time frame after collection.

Swabs are not collected from the site of infection.

3.9 REQUESTING ADDITIONAL EXAMINATIONS

Additional tests may be added to the original request by contacting the laboratory and submitting a

completed request form.

Any additional Test must be requested within 1 week from the date of sample collection. MBG Lab is

usually able to perform further tests on stored samples as long as the additional request is made within the

appropriate timeframe. However, our ability to produce a valid result from the add-on test will be

dependent on the availability of the appropriate sample, the stability of the analyte, how the sample has

been stored, and the remaining sample volume.

4 RESULTS

The main method of reporting results originating from this lab is by the production of an electronic copy of

the signed report (encrypted emails). Original reports, once signed by authorized personnel, are available

for collection from the Lab. An electronic copy of all these reports is held on the Laboratory Information

Management System (LIMS).

The results shall be sent to the destination (person) specified in the request form and by the means

selected (encrypted e-mail, fax, and telephone or through post/courier). If this section of the request form

was not completed, the results shall be made available as a hard copy only to the requester. If a person

other than the requester (E.g. hospital driver) picks up the result from MBG Lab, it is the responsibility of

the person delivering the report to ensure that the report reaches safely to the authorized person. Such

reports are provided in a sealed envelope to ensure confidentiality of the client.

Any interpretations of the results, if required, are clearly mentioned in the report. MBG Lab offers no

further interpretation.

5 REPORTING EMERGENCY/CRITICAL OR URGENT RESULTS

In case of urgent or emergency/critical requests, the results are bought to the immediate attention of the

authorized requesting person to prevent situations where there is potential of danger or harm to the

patient. This is usually done by calling the authorized person using the contact number provided on the

request form.

A copy of the original report may also be sent through an encrypted e-mail or fax. The original report is

available at the Lab for collection.

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6 FEEDBACK

We aim to provide a high quality service and are always aiming to make improvements to the services we

offer. In order to do this we value comments from our users. If you have any comments please feel free to

contact our Quality Management Team. Similarly, acustomer feedback form is available on our website

www.mgb.ae .

7 COMPLAINTS

Users of the services can contact the lab through fax, email or telephone to log any complaints or

grievances that may occur (refer Section 1.3 Contact Information). These will be acknowledged and a

written response will be given to the plaintiff concerning the investigations and actions taken by the

laboratory to rectify the issue. All complaints are monitored and if there is any, this is discussed during

weekly lab meeting. A customer feedback form is available on our website www.mgb.ae.

8 CUSTOMER SATISFACTION

We are continually improving our services and value the opinion of our users in order to provide a service

that reflects the highest of standards while also being user friendly. We issue our users with ‘Customer

feedback form’ for completion. We would encourage you to partake in these surveys so that our services

can reflect your views. We also welcome any feedback you have regarding the services at any time by

communication through fax, email or telephone (refer Section 1.3 Contact Information). A customer

feedback form is available on our website www.mgb.ae.

9 VETERINARY MOLECULAR DIAGNOSTIC SERVICES

9.1 Pathogen Identification Assays

MBG Lab offers several different assays to detect various pathogenic bacteria, viruses and parasites from

different biological samples. The vast Majority of our Pathogen ID assays are accredited according to ISO

17025:2005.

9.2 Avian Sexing

DNA based avian sexing is the method of choice for bird owners worldwide. This is determined by

amplifying specific genes on the sex chromosomes. A range of different techniques are utilized for different

avian species all of which have been accredited to ISo17025: 2005.

9.3 Animal Genotyping

Breeders are often required to prove the true identity of an animal through a verified genotyping

certificate. At MBG Lab, this is done using microsatellite markers that are used to genotype different

animals such as equine, camel, bovine, ovine, falcon, dolphin, tiger etc. Our Genotyping service has been

ranked 1 by ISAG for a range of animals including Camel, lama, Equine.

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9.4 Paternity Verification

Accurate pedigree information is important for animal breeders and is often required to resolve paternity

of disputed cases. Currently, MBG Lab provides Genotyping and Parentage Verification assays for Equine

and Camelids.We have two panels of markers developed in-house and ranked 1 by ISAG allowing MBG to

accurately verify parents of both species.

9.5 Species identification

MBG Lab has developed a real-time PCR multiplex assay accredited to 17025:2005 that identifies and

quantifies minute amounts of mixed-species nuclear DNA of cattle (Bostaurus), horse (Equuscaballus), goat

(Capra hircus)/sheep (Ovisaries), pig (Sus scrofa)and camel (Camelusdromedarius) in a species-specific

manner. The assay’s ability to accurately detect and identify target DNA in multi-species mixtures is crucial

when the target specimen may be overwhelmed by non-target specimen.

9.6 Avian Species Differentiation

MBG Lab has developed a test to different species of avian specifically chicken and houbara. This test is

currently offered for research purposes only.

9.7 Equine Coat Colour

Horse coat colour genetics is important for offspring selection by horse breeders. Several Single Nucleotide

Polymorphism’s (SNP’s) as well as mutations in genes linked to coat colour are analysed at MBG Lab to

ascertain specific coat colour phenotype.

10 MEDICAL MOLECULAR DIAGNOSTIC SERVICES

Pathogen Identification Assays

MBG Lab offers several different assays to detect various pathogenic bacteria, viruses and parasites from

different biological samples. We provide a fully accredited service ISO15189:2012 for the detection of

H1N1, Influenza, MERS Corona and Zaire Ebola testing.

MBG Lab is the first centre to carry out H1N1 testing, through Real-Time PCR, in the UAE. Since May 2009,

we have tested over 10,000 samples and are among one of the few centres in Dubai with the essential

technology and expertise to provide H1N1 diagnostic services.MBG Lab is currently the only accredited

centre in UAE to carry out Zaire Ebola testing.

We also provide fully validated assays for the detection of a number of other pathogens including zika virus

and viral meningitis.

11 CYTOGENETICS

Cytogenetics is a dynamic field both in technological development and clinical applications and involves

conducting various types of chromosomal analyses. At MBG lab, chromosomal analysis is done for variety

of species such as Human, Equine, Camelids, Avian, etc. Techniques such as standard karyotyping together

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with Fluorescent In Situ Hybridization (FISH) are used to investigate gains or losses in genetic material that

help in the identification and characterization of genetic abnormalities in different species using blood or

tissue samples.

The type of technique used for investigation depends on the specificities of each case. Each case is dealt

with according to its list of conditions such as:

Multiple congenital abnormalities

Gonadal dysgenesis

Birth defect

Infertility: chromosomal analysis is recommended for cases with a history of infertility, more than

two spontaneous abortions, abnormal gestational development, fetal death, or family history of

genetic abnormalities.

11.1 Classical Karyotyping

This is done using the facilities of Olympus microscope and the Applied Imaging software. Chromosomal

karyotyping helps in detection of any abnormality present at the chromosomal level.

11.2 Fluorescent in Situ Hybridization

FISH is considered a cytogenetic as well as molecular cytogenetic technique. It

depends on the use of a nucleic acid probe that hybridizes to the entire chromosome

or a single unique DNA sequence. FISH helps in the detection of gains or losses in

DNA.

11.3 Sperm Viability Test

Fertility is an important aspect for animal breeders and consequently has high

economic impact. Male fertility, however, cannot be accurately assessed using the absolute assay and

traditional semen analysis. Therefore, semen samples need to be subjected to different tests which can

improve the prediction of the fertility status.

Simultaneous evaluation of head and tail viability, acrosome integrity and mitochondrial function of

spermatozoa permit an accurate evaluation of sperm quality and fertility status.

12 REGENERATIVE STEM CELL THERAPY

12.1 MBG-Stem

What is MBG-Stem Regenerative Cells and what are their benefits?

MBG-Stem Regenerative cells are your animal’s own natural healing cells that help

regenerate injured tissues. These cells are isolated from the animal’s adipose (fat)

tissue and consist of stem cells, progenitor cells and other bioactive cells which enable

a more natural healing process.

The benefits of using MBG-Stem Regenerative Cells are that:

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These cells can turn into various types such as tendon, ligament, bone, cartilage etc.

They promote proper tissue remodelling, thus, preventing the formation of scar tissue.

They produce growth factors that help promote regeneration and reduce inflammation.

There is only 48 hour turn-around time as there is no initial culturing required.

Stem cells are your animal’s own naturally occurring cells.

Fat is a non-vital, renewable tissue source providing a higher yield of regenerative cells.

Lacks political/ ethical controversy, unlike embryonic stem cells.

Easy and safe to collect.

What is the procedure?

Is it safe?

This technology is in accordance with FDA and USDA guidelines.

Auto- immune tissue reaction is minimal as the animal’s own cells used.

Minimally manipulated cells are not differentiated or modified.

Stringent processing protocols and Quality Control methods.

Peer reviewed publications support the safety of autologous cell therapy.

Very low reported adverse events.

Current uses MBG-Stem cell therapy can be used to treat large animals such as horse, camel, etc. as well as small

animals such as dogs, cats, etc. A number of conditions are treated, which include:

Osteoarthritis – chronic

Orthopedic – soft tissue

o Tendons

o Ligaments

o Fractures

12.2 Platelet – Rich Plasma

Another service provided under MBG-Stem is Platelet-rich Plasma or PRP. This is an exceptional way to

treat joints and other injuries and interestingly it can be administered to the injured animal in conjunction

with Stem cell therapy for a better outcome.

Step 1: Fat collection – A small sample of fat is collected from the animal by the veterinarian and shipped to MBG Lab.

Step 3: Injection – The Regenerative Cells are injected back into the animal by the veterinarian.

Step 2: Processing – The Regenerative Cells are isolated from the fat tissue at MBG Lab and shipped to the veterinarian.

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12.3 Banking

As a part of the MBG-Stem service, we also provide storage of the regenerative cells for future use. After

following the procedure outlined above, part or all of the cells may be banked at owner's request for

future need. This offers the owner and/or veterinarian the option to preserve their animal’s stem cells for

long-term storage in liquid nitrogen, in case the animal needs to be treated for the existing injury or for a

new injury.

13 MOLECULAR GENOMICTECHNIQUES

MBG Lab carries out a variety of molecular genetics techniques for both research and diagnostic purposes.

Some of the services we offer include:

13.1 STR Analysis

Microsatellites markers can be used for:

Individual identification

Parentage verification

Population and Linkage analysis

Investigation of forensic cases

The microsatellites are identified using the Applied

Biosystems 3730/3730xl DNA Analyzer which is an

automated, high-throughput, capillary electrophoresis

system used for analyzing fluorescently labeled DNA

fragments. These fragments are then analyzed using the

Genemapper® software, a flexible genotyping software

package that provides DNA sizing and quality allele calls for

all Applied Biosystems electrophoresis-based genotyping

systems.

13.2 Sequencing

DNA sequencing is the process of determining the nucleotide order of a given DNA fragment, based on the

dideoxy method or Sanger method. This is the gold standard

for sequencing and a confirmatory tool for:

Microbial identification

Species identification and differentiation by comparing

mitochondrial sequences

SNP identification

Screening physically mapped sequence tagged sites

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(STS).

At MBG Lab this is done using the 3730xlAnalyzer from Applied Biosystems.

13.3 SNP Genotyping

Genotyping of SNPs is extremely important as SNPs can be associated

with different genetic traits, susceptibility to diseases or responses to

drug therapies. SNP genotyping facilitates numerous association studies

of complex genetic traits.MBG Lab houses the state-of-the art, high

resolution iScan® system from Illumina to carry out SNP genotyping

studies. This enables rapid, sensitive and accurate imaging of Illumina

array based genetic analysis products. SNP genotyping at MBG Lab include methods such as:

Genome re-sequencing for SNP discovery, confirmation and genotyping.

Genotyping using custom made SNP array which uses thousands of characterized SNPs on a

microchip that enables the selection of specific traits in an individual.

13.4 Gene Expression Profiling

Gene expression profiling is the measurement of the activity (the expression) of thousands of

genes at once, to create a global picture of cellular function.

These profiles can, for example, show how the cells react to a

particular treatment or stress conditions.

MBG Lab has the Q Array2 microarray printing machine

(Genetix) and a GenePix scanner (Axon Instruments) which are

particularly useful for in-house microarray printing and

scanning.

13.5 Construction of genomic and cDNA libraries

At MBG Lab, genomic, SSH and cDNA libraries have been created for

different species using the picking and subclone management system,

QPIX 2, (Genetix). This machine enables tasks such as picking colonies

(e.g. E. coli, yeast, phage, etc), replicating plates/libraries, setting up

screens, as well as cherry picking wells of interest. Further applications

include rearraying (hit picking), Y2H, SGA, gridding and phage display.

14 RESEARCH AND DEVELOPMENT

MBG Lab’s research theme include the areas of diagnostics and genomics covering genotype & phenotype

association studies, mitochondrial genome, gene regulation and expression, infectious diseases, functional

genomics, fertility studies, developing species specific probes and chromosome karyotyping. MBG Lab is

well equipped with various high throughput liquid handlers for DNA & RNA extraction, sequencing

machines, robotic colony pickers, high throughput microarray machines, and real-time PCR machines.

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Due to the excellent core facility offered by MBG Lab, the department is also committed to train and

educate the next generation of students and scientists. MBG Lab provides a full spectrum of research

training opportunities with the underlying common theme of Molecular Genetics. We have ongoing

research projects at the Graduate and Postgraduate levels with students currently registered with various

universities, both national and international.

Some of the projects undertaken so far at MBG Lab include:

Evaluation of Microsatellite Markers from various Camelids in dromedary camel

Creation of Microsatellite Marker panels for Genotyping and Parentage Verification assay in

Camelids.

Construction and comparison of Chromosomal karyotype of several Camelids species and their

Hybrids

Complete Sequencing of Mitochondrial DNA of Dromedary Camel

Sequencing several Equine Genes

Identification of unique SNPs in equine genome

Please do not hesitate to contact us for further details and potential collaborations.

15 EDUCATION AND TRAINING

Being one of the best-equipped genetic laboratories in the country, we are proud to offer technical and

hands-on courses for various molecular biology applications. These courses are given in a long or short

form depending on the choice of the students. A certificate of completion will be furnished upon

completion of training. We also support research students in their projects.

The six different core molecular genetics courses are:

a- Nucleic acid extraction

b- PCR, Real time PCR and Gel Electrophoresis

c- Sequencing

d- Cytogenetics and FISH

e- Microarray

f- Genotyping

Kindly refer to MBG Website for more details on the courses.

molecular biology and genetics …however, urgent laboratory investigations are undertaken when...

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