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An Advanced cDNA Synthesis Kit for RT-qPCR: High Capacity and Wide Dynamic Range
Advantages of a High-Capacity cDNA Reverse Transcription Kit
A high-capacity cDNA reverse transcription kit can provide significant savings, efficiency, and reproducibility for laboratories that use substantial quantities of cDNA for protocols such as screening and microarrays. The iScript™ advanced cDNA synthesis kit has a capacity of up to 7.5 μg of RNA per 20 μl reaction — over three times the capacity of other kits.
Benefits of using a high-capacity reverse transcription kit include:
- Increase your reverse transcription throughput
- Fewer reactions save time and money
- Analyze more targets from fewer reactions
- Reduce the effects of inter-assay variability
- Improve reproducibility
- Detect low level target genes
- Master mix reduces manipulations and reduces pipetting error
- Low reaction volume with high concentrations of RNA
- Extended dynamic range
Choosing a cDNA Reverse Transcription Kit
When choosing a kit for high-capacity cDNA reverse transcription, in addition to a high yield of first-strand cDNA product, there are several other considerations.
Does the kit provide a high yield of full-length cDNA and avoid 3' and 5' bias? The iScript advanced cDNA synthesis kit uses priming with a combination of oligo(dT) and random primers which has been shown to increase the number of full-length cDNAs. Using oligo(dT) alone gives 3' bias and increases the population of transcripts that terminate prematurely if there is a region of strong secondary structure. Random primers alone, due to internal priming, tend to give shorter transcripts, which can lead to an overestimation of copy number in real-time PCR. An optimized combination of oligo(dT) and random primers will provide more full-length cDNA and less bias than either type of priming alone.
Do you want the convenience and increased accuracy of a master mix? The majority of the available high-capacity cDNA reverse transcription kits have all the components in separate tubes. Our enhanced formulation provides all the components you need in just two tubes for both convenience and flexibility. This significantly reduces the pipetting required when compared to other kits but also provides some flexibility.
How much input RNA do you need to transcribe? How high a capacity is optimal for your downstream analyses? For multiplex PCR assays and screening total RNA from cells and tissues, a large quantity of cDNA is often required. RNA quantity can be an important consideration when multiplexing quantitative PCR or when targets include low-abundance RNAs.
Summary
The ability to process a wide range of input RNA concentrations is an advantage of a high-capacity cDNA reverse transcriptase kit, as it increases its flexibility. Another major benefit of using a high-capacity kit is the reduction in inter-assay variability and subsequent improved reproducibility, which is particularly important for accurate detection of low-abundance RNAs and for large multiplex assays. Finally, there is increased throughput and a saving of time, reagents, and precious samples. Try a high-capacity cDNA reverse transcription from Bio-Rad to see the difference in throughput and accuracy.