RT2 First Strand Kit
For cDNA synthesis and genomic DNA elimination in RNA samples for use with RT2 Profiler PCR Arrays and RT2 lncRNA PCR Arrays
Products
The RT2 First Strand Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
RT2 First Strand Kit (12)
Cat. No. / ID: 330401
For twelve 20 µl first strand cDNA synthesis reactions; Buffer GE (24 µl), 5x Buffer BC3 (48 µl), RE3 Reverse Transcriptase Mix (24 µl), Control P2 (12 µl), Nuclease-Free Water (1 ml)
RT2 First Strand Kit (50)
Cat. No. / ID: 330404
For fifty 20 µl first strand cDNA synthesis reactions; Buffer GE (100 µl), 5x Buffer BC3 (200 µl), RE3 Reverse Transcriptase Mix (100 µl), Control P2 (50 µl), Nuclease-Free Water (1 ml)
Features
- Rapid and efficient first-strand cDNA synthesis
- Complete elimination of genomic DNA
- Results with as little as 25 ng total RNA per reaction
- cDNA immediately ready for real-time PCR
- External RNA control to monitor enzyme inhibitors
Product Details
The RT2 First Strand Kit provides a rapid and convenient procedure for efficient first-strand cDNA synthesis and genomic DNA elimination in RNA samples. The synthesized cDNA is ready to use in real-time PCR expression analysis of multiple genes.
Principle
The RT2 First Strand Kit provides a rapid and convenient procedure for efficient cDNA synthesis from mRNA for use in gene expression assays.
The kit includes a proprietary procedure to effectively eliminate contaminating genomic DNA from RNA samples before reverse transcription. Random hexamers and oligo-dT prime reverse transcription in an unbiased manner, and a reverse transcriptase synthesizes cDNA product with optimal yield and length. A built-in external RNA control helps monitor reverse transcription efficiency and test for enzyme inhibitors. The RT2 First Strand Kits are optimized for real-time PCR-based gene expression analysis using RT2 Profiler PCR Arrays and RT2 qPCR Primer Assays, and can be successfully used with other commercially available gene expression analysis assays.
Procedure
The proprietary Buffer GE and sample are mixed and then incubated on a thermal cycler. After addition of the enzyme and additional buffer, the sample is then reverse transcribed using any thermal cycler. The transcribed cDNA is immediately ready for use in downstream applications. We recommend RT² Profiler PCR Arrays or RT² qPCR Primer Assays for further analysis.
RT² First Strand Kits yield results with as little as 25 ng or as much as 5 µg total RNA per reaction. However, the optimal amount of starting material depends on the relative abundance of the transcripts of interest. Lower abundance transcripts require more RNA; higher abundance transcripts require less RNA. Greater amounts of input RNA yield greater number of positive calls (i.e., gene expression detection) in the linear dynamic range of the method.
Applications
cDNA synthesized using the RT2 First Strand Kit yields excellent results in real-time PCR expression analysis of multiple genes.
Resources
Safety Data Sheets (1)
Kit Handbooks (3)
Brochures & Guides (1)
Application Notes (1)
Instrument Technical Documents (1)
Certificates of Analysis (1)
FAQ
What do I need to complete a RT² qPCR Primer Assay?
What is the recommended solution in which to store RNA samples that will be used as templates for cDNA synthesis?
How can I avoid or remove genomic DNA contamination from the total RNA preparation?