These easy-to-use and highly efficient kits are designed for the synthesis of first-strand cDNA using your choice of cDNA priming method
- Separate components for RT priming method, (oligo-dT, random hexamer, or GSP)
- Highly sensitive first-strand cDNA synthesis of RNA sequences up tp 12 kb for
- Broad linear dynamic detection range with limiting (10 pg) or plentiful (4 µg) total RNA samples
qScript™ reverse transcriptase is a mixture of an engineered RNase H+ MMLV-RT and recombinant ribonuclease inhibitor protein that has been specifically optimized for use with the provided qScript™ flex reaction mix. This unique 5x master mix of buffer, magnesium, stabilizers, and dNTPs simplifies reaction assembly and ensures robust and reproducible synthesis of first strand product from 10pg to 2µg of total RNA or purified polyA+ RNA template.
The resulting cDNA product is directly compatible with current real-time RT-PCR methods or end-point RT-PCR. Length of cDNA product is dependent upon priming strategy and quality of the RNA template. Oligo dT or GSP can be used for long RT-PCR of RNA targets up to 12 kb. Random primer is suitable for RT-PCR of RNA targets less than 1kb. Additionally, the location of the target sequence in the mRNA should be considered when selecting a priming method. Oligo dT primer is the preferred choice for RT-PCR of 3' end sequences. The efficacy of RT-PCR of 5' end regions is often improved by using random or gene-specific primers.
The kits provide optimized reagents for priming with oligo(dT)20, random primer, gene-specific primer (GSP), or any combination thereof. A key component is a proprietary enhancer compound that improves cDNA priming efficiency. This is provided as both a separate solution for use with your gene-specific primers, and pre-blended with the supplied Oligo dT or Random Primer solutions.
Ordering information: Any of the three priming methods is suitable when these kits are used for qRT-PCR. However, it is recommended that a mixture of oligo dT and random primer be used to achieve consistent and unbiased first-strand synthesis over a broad range of RNA template concentrations. If more stramlined procedures are preferred, see either qScript™ cDNA SuperMix kits (101414-102 series) or cScript cDNA synthesis kits (101414-096 series) for optimized gene expression procedures.
