Description
One-Step RT-PCR Kit
One-Step RT-PCR Kit is a convenient and quick solution for performing RT-PCR in a single step. RT-PCR is a well-used technique to amplify double-stranded DNA fragments from RNA templates. In the first reverse transcription step, the reverse transcriptase synthesizes cDNA based on the RNA template. In the second PCR step, Taq DNA Polymerase synthesizes DNA using the synthesized cDNA as template.
The One-Step RT-PCR Kit combines reverse transcription and PCR into one reaction step, which eliminates the need for two-step reactions. The one-step strategy avoids the transfer of reaction mixtures from the reverse transcription step to the PCR step, and reduces the contamination risk and minimizes tedious sample handling.
The unique formulation of both buffer and enzyme mix delivers reliable and consistent results. The optimized buffer components allow for efficient reverse transcription and PCR with secondary structure and ensure complete cDNA synthesis and specificity of the PCR fragments.
One-Step RT-PCR Kit data. The kit was used in two samples with different amplicon sizes.
The buffer of the One-Step RT-PCR Kit includes dNTPs, MgCl2, PCR enhancer, and PCR stabilizer. The enzyme mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, enzyme enhancer, and enzyme stabilizer. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize DNA strands using RNA as template. The source of the Reverse Transcriptase is a cloned reverse transcriptase gene from M-MuLV. Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a low 5´→ 3´ exonuclease activity. The source of the Taq DNA Polymerase is a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1.
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Features
- cDNA synthesis coupled with PCR in one step
- Robust and reliable reactions
- Unique buffer and enzyme formulations generate consistent results
- PCR fragments have 3′-dA overhangs
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