Reduce your reverse transcription time to 10 minutes using the new Transcriptor Universal cDNA Master. This optimized, ready-to-use reaction mix is the convenient, fast solution for cDNA synthesis in two-step real-time RT-PCR (see Figure 1). Transcriptor Universal cDNA Master is compatible with LightCycler® Instruments and all other real-time instruments requiring normalization with a reference dye.

J Achieve high sensitivity with all, even the most difficult templates.

Reverse transcribe RNA with high secondary structures at temperatures up to 60°C.

J Reliably quantify your qRT-PCR experiments. Obtain consistent results over a linear dynamic range of at least six orders of magnitude.

J Increase two-step qRT-PCR sensitivity and specificity. Produce lower cycle threshold (Cq) values, starting with

target amounts as low as 0.1 pg of total RNA.

J Reduce setup time. Just add RNA – two vials contain all necessary reagents, including random hexamer primers, nucleotides, buffers, and enzymes.

Transcriptor Universal cDNA Master Faster cDNA Synthesis – On Any RealTime-PCR Instrument

Figure 1: Transcriptor Universal cDNA Master, in combination with the RealTime ready Cell Lysis Kit, produces much faster time-to-result than conventional workflows.

Convenience

30 60 90 120 150

ConventionalWorkflow

AcceleratedWorkflow

Time-to-Result (min)

RNA Preparation

cDNA Synthesis

Real-Time PCR

For life science research only.Not for use in diagnostic procedures.

Figure 3A: Use Roche products for your gene expression workflow, from cell monitoring to relative quantification.

Published by

Roche Diagnostics GmbHRoche Applied ScienceWerk Penzberg82372 PenzbergGermany© 2010 Roche Diagnostics GmbHAll rights reserved.

06418155001 0810

Performance and Convenience – The Perfect Combination

Choosing the correct enzyme for reverse transcription and cDNA synthesis is critical for obtaining high yields of quality cDNA. Highly thermostable Transcriptor Reverse Transcriptase has a broad temperature range and is, when combined with the supplied buffer, ideal for high-temperature reverse transcription. Excellent performance and linear response in real-time RT-PCR is ensured for both low- or high-copy transcripts.

Combine the Transcriptor Universal cDNA Master with the RealTime ready Cell Lysis Kit to accelerate real-time PCR analysis of cell lysates (see Figure 3A).

B C

Figure 3B: The effect of doxorubicin, a drug used for cancer treat-ment, on the proliferation and viability of HepG2 cells. A human liver carcinoma cell line was analyzed using the xCELLigence System in a 96-well E-Plate. Figure 3B shows that with doxorubicin concentrations between 2 µM to 200 µM, a signifi cant reduction in cell growth is observed 8 hours after treatment. Cells were lysed using the new RealTime ready Cell Lysis Kit. RNA-containing cell lysate was reverse transcribed using the Transcriptor Universal cDNA Master. Resulting cDNA was amplifi ed with LightCycler® DNA Master Probes and the RealTime ready Human Apoptosis Panel using the LightCycler® 480 Instrument.

contr200 nM

2 µM20 µM

200 µM

Roche Applied Science provides comprehensive reverse transcription products. For more information, visit http://www.cDNA.roche.com

Product Pack Size Catalog Number

Transcriptor Universal cDNA Master

100 x 20 µl reactions 05 893 151 001

RealTime ready Cell Lysis Kit

500 reactions 05 943 523 001

Ordering Information

Figure 3C: Transcript levels of various genes at different doxorubicin treatment levels (Figure 3B) were quantifi ed by real-time PCR. For each transcript, the average of triplicates was color-coded and plotted in a heat map using the Gene Pattern Software of the Broad Institute (http://www.broadinstitute.org/cancer/software/genepattern/index.html).

CP-value color conversion

Housekeeping Genes

RealTime cell analysis using the xCELLigence System

Rapid sample preparation using the RealTime ready Cell Lysis Kit

Reverse Transcription using the Transcriptor Universal cDNA Master

RealTime PCR using the LightCycler® 480 System and RealTime ready Apoptosis Panels

Figure 2: Transcriptor Universal cDNA Master produces earlier Cqs and higher sensitivity using real-time PCR compared to the reverse transcriptase kit from supplier B.

For life science research only. Not for use in diagnostic procedures.LIGHTCYCLER, REALTIME READY and XCELLIGENCE are trademarks of Roche. E-PLATE and ACEA BIOSCIENCES are registered trademarks of ACEA Biosciences, Inc. in the US.

Gene expression heatmap of Doxorubicin treated cells

56.4

52.8

48.3

43.8

39.3

34.8

30.3

25.8

21.3

16.8

12.3

7.8

3.3

Cycles

Amplification Curves

Flu

ore

scen

ce (

46

5-5

10)

2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44

Transcriptor Universal cDNA Master

Supplier B

A

Time (in Hour)

No

rmal

ized

Cel

l In

dex

3.50

3.15

2.80

2.45

2.10

1.75

1.40

1.05

0.70

0.35

0.00 0.0 16.0 32.0 48.0 64.0 80.0 96.0