REVERSE TRANSCRIPTION PCR
Reverse Transcription PCR (RT-PCR) can be performed as a One-Step or Two-Step reactions. Two-Step RT-PCR is useful for detecting multiple messages from a single RNA sample or when you need maximum flexibility in selecting PCR conditions.
The One-Step RT-PCR method offers the capability of performing both cDNA synthesis and PCR amplification in a single tube which prevents contamination and promotes rapid processing for a large number of samples. Since the entire cDNA is amplified, One-Step RT-PCR can also provide greater sensitivity. Table 1 compares the benefits of One-Step and Two-Step RT-PCR methods.
Biotools offers RT-PCR kits with high transcriptional efficiency, as well as high yield, sensitivity and specificity for customer RT-PCR needs, including end-point and real time quantitative RT-PCR detection kits.
High-fidelity DNA polymerases and reverse transcriptases are valuable for minimising the introduction of errors in amplified products. The use of high-fidelity amplification conditions is essential when analysing small amounts of template DNA or rare molecules in heterogeneous samples.
Almost all Biotools RT-PCR kits have been developed with a new thermostable retrotranscriptase that offers advantages with respect to other enzymes available elsewhere (See Figure):
- Robust cDNA synthesis across a broad range of temperatures from 37 to 77 °C
- Excellent cDNA yields
- Ability to reverse transcribe through a secondary structure
- High stability
High Sensitivity in qRT-PCR
Biotools Reverse Transcriptase was more sensitive than MMLV Reverse Transcriptase, delivering an earlier Ct value.
TABLE 1: Download Comparison of One-Step and Two-Step RT-PCR Procedures
TABLE 2: Designed to help in the selection of Biotools RT-PCR Kits
Related Products
- dNTPs
- Primers and Probes
- Fluorophores and Dyes
- PCR Enhancer
- Amplification Controls
- Buffers and Solutions
- DNA and RNA Purification
- Electrophoresis Reagents
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RetroTools® TwoStep kit