pcr- restriction fragment length polymorphism (rflp)...q1_student fatima and hanan author omar harb...
Post on 03-Mar-2021
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PCR- Restrictionfragmentlengthpolymorphism(RFLP)
Students’presentation1By
FatimaZahran &Hanan Mahmoud
Principleofthetest
• PCR-RFLP is a techniquethat combines the PCRamplification with thedigestion of PCR productsby restriction enzymes(RE).
• Cleaving DNA intofragments of certainsizes, whose analysis onagarose gel results indifferent patterns offragment sizes, enablingthe identification.
First:Leishmania PCRanalysis
DNAextractionfrombloodsamples
ITS1PCRassay
Forspeciesidentification
PCRproductpurification(Qiagene kit)
RFLPusing
Hae IIIenzyme
M LT LM M LT LM
Buttherearesomedisadvantagesforthistechnique
• It does not detect all sequence and lengthvariations within or among amplicons, sincethe restriction enzymes used only recognise asmall number of potentially variable sites.
• Additionally, it requires large amount ofhighly pure DNA
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