High Efficient Reverse Transcription Kit for Real-time PCR

After reverse transcription, the RT products were diluted with an equal volume of dH2O, and 2 µl was used for real-time PCR (corresponds to 50 ng total RNA/PCR reaction). Custom TaqMan assays were used to analyze expression of GAPDH and RNase P using an ABI 7900HT Fast Real Time PCR System. All reactions were performed in duplicate. The PCR conditions were 95°C for 10 min, then 40 cycles of 95°C for 15 sec and 55°C for 1 min. Data was analyzed using SDS v2.4 software automated analysis.

Table I. Ct values obtained from real-time PCR amplification.

* All reactions were run on an Applied Biosystems 7500 Fast Real-Time PCR system.

Cultured cell lysis & cDNA Synthesis kit for real-time PCR

For the 0.2 µmol synthesis scale, monomer coupling is done at a 40-50-fold excess. Doing so for larger scale synthesis (such as for a 1.0 µmol scale) would be cost-prohibitive. Large-scale syntheses are done only at 10-fold mole excess of amidites. To increase the final yields of these large-scale synthesis, the coupling times are extended, so to increase coupling efficiencies.

Modifications for Real-time PCR

For the 0.2 µmol synthesis scale, monomer coupling is done at a 40-50-fold excess. Doing so for larger scale synthesis (such as for a 1.0 µmol scale) would be cost-prohibitive. Large-scale syntheses are done only at 10-fold mole excess of amidites. To increase the final yields of these large-scale synthesis, the coupling times are extended, so to increase coupling efficiencies.

Real-Time PCR Primer Design / Oligo Synthesis | …

The program is a comprehensive real time PCR primer and ..

Available for both SYBR® Green and TaqMan detection, One-Step PrimeScript RT-PCR Kits allow all RT-PCR steps to be performed in a single tube, simplifying reaction set-up, minimizing the risk of contamination, and, with a reverse transcription step of just 5 minutes, allowing rapid analysis. In Figure 2, the was used to analyze HPRT1 expression in human liver total RNA (2 pg–2 µg) in a single-tube reaction (cDNA synthesis and real-time PCR performed in tandem). This one-step protocol using PrimeScript RTase allowed high sensitivity and specificity detection of HPRT1 mRNA across a wide range of template RNA concentrations (Figure 2).

Frequently asked questions for Oligo Synthesis and siRNA

Ordinarily, it can take 30 to 60 minutes to synthesize cDNA templates for real-time RT-PCR (reverse-transcription PCR). With PrimeScript Reverse Transcriptase, RT reactions can be completed in as few as 15 minutes. Not fast enough? Use a one-step PrimeScript kit and set your reverse transcription step to just 5 minutes.

What should I use for cDNA synthesis? - ResearchGate

What should I use for cDNA synthesis

purification is standard (no additional charge) for all our dual-labeled probes and/or multi-labeled oligos, as well as for all our RNA and large scale oligos.

All Real-Time Ready cDNA Synthesis Products

for real-time PCR and with a ..

For oligos of standard length (≤ 33 nucleotides) we guarantee a certain yield of DNA (OD260 applying to a 20-mer) to be delivered according to the respective scale of synthesis and mode of purification.

for real-time PCR, random primers ..

Real-time qPCR Probes: Oligos for real-time PCR, ..

Reduce reverse transcription reaction time from >60 minutes to 15 minutes and generate the same reliable real-time PCR results. The is optimized for fast, efficient synthesis of cDNA templates for real-time PCR.