First Strand cDNA Synthesis BioGenomics™ Kit is developed based on cDNA synthesis protocol. The kit generates full-length first strand cDNA that is ready for PCR amplification. An easy to follow protocol is provided. The kit contains sufficient reagents for 100 cDNA synthesis reactions.
Intended Use
Immediate PCR amplification of known genes Comparison of gene expression patterns between biological samples Gene mutation analysis Gene cloning and target sequencing
Test Principle
This First Strand cDNA Synthesis BioGenomics™ Kit provides all necessary components to perform 100 first strand cDNA synthesis reactions from total RNA or poly(A) RNA. The system uses Moloney Murine Leukemia Virus Reverse Transcriptase with weak activity of RNase H that allows the synthesis of full-length cDNA from long templates. Oligo(dT)18 and random hexamer primers are provided. Specific primers can also be used with the kit.
The cDNA synthesized using this system can be used directly in PCR amplification. The kit is optimized for maximum yield of full-length cDNA. The addition of ribonuclease inhibitor lowers the risk of mRNA degradation during the reaction.
Quality Control
All kit components are DNase-, RNase-, and protease-free. Each component has been tested for purity and efficacy in first strand cDNA synthesis.
Kit Components
MMLV reverse transcriptase, 100unit/ul, 1x100ul Reaction Buffer, 5x, 1x1ml Oligo(dT)18, 0.5ug/ul, 1x100ul Random Hexamers, 0.2ug/ul, 1x100ul RNasein, 10units/ul, 1x100ul dNTP, 10mM each, 1x250ul Human Placenta total RNA, 1.25ug/ul, 1x10ul Upstream Primer, 25um, 1x50ul Downstream Primer, 25um, 1x50ul DEPC H2O, 1x1.25ml
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.