It is often necessary to obtain a large quantity of nucleic acid to study or detect individual genes or specific DNA regions or mutations of interest. Polymerase Chain Reaction (PCR) is the most used DNA amplification method in molecular biology. For experiments where detection and quantification are required instead of isolation, quantitative PCR (qPCR) uses real-time fluorescence to measure the amount of a DNA target present at each cycle during a PCR. Use of hydrolysis probes such as TaqMan® or a double-stranded DNA binding dye such as SYBR® Green are the most common methods of generating a fluorescent signal.
RNA molecules can also be detected via the use of Reverse Transcriptase (RT), which is an RNA-dependent DNA Polymerase. RT polymerizes a strand of DNA that is complimentary to the original RNA template and is referred to as cDNA. This cDNA can be further amplified through PCR, qPCR or detected in a single reaction using one-step RT-qPCR.
G-biosciences offers a wide selection of enzymes and kits useful for various PCR, qPCR and RT-qPCR. Some of our proprietary products are listed below.
- Taq DNA Polymerase possesses a 5´→3´ polymerase activity and a 5´-flap endonuclease activity that can amplify DNA up to 5kb with an elongation velocity of 0.9-1.2kb/min at 70-75°C.
- Reverse Transcriptase (RT) is a recombinant Moloney Murine Leukemia Virus (MMLV) reverse transcriptase with reduced RNase H activity. This RT is an RNA-directed DNA polymerase which lacks 3´→ 5´ exonuclease activity and can produce full-length cDNA from long RNA templates.
- RNase Inhibitor (Murine) is a recombinant protein of murine origin which specifically inhibits RNases A, B and C. Moreover, it does not inhibit polymerase activity when mix with Taq DNA Polymerase and Reverse Transcriptase.
- First strand cDNA synthesis kit is in a 5X master mix form which contains Reverse Transcriptase (RT), recombinant RNase inhibitor, dNTPs and an optimized buffer with protein stabilizers.
- SYBR® Green based qPCR 2x master mix contains chemically modified hot start Taq DNA polymerase, SYBR® Green I fluorescent dye, low ROX reference dye, dNTPs and optimized buffer with stabilizers.
- Probe-based qPCR 2x master mix contains Taq DNA polymerase, low ROX reference dye, dNTPs and optimized buffer with stabilizers.
- Probe-based one step RT-qPCR 2x master mix contains Reverse Transcriptase, Taq DNA Polymerase, low ROX reference dye, dNTPs and optimized buffer with stabilizers. This master mix is ideal for high-throughput real-time PCR screening and validation which requires minimal handling and offer robust and consistent results.