BcaBEST™ RNA PCR Kit
Applications:
- Single-tube RT-PCR, using LA PCR technology for better PCR performance
- Reverse transcription of RNAs with high amounts of secondary structure
Features:
- Any RNA can be used as a template. Especially useful for GC rich RNA or RNA with secondary structure.
- Can amplify messages up to 5 kb in length.
- BcaBEST™ DNA Polymerase is utilized to carry out transcription at an optimum reaction temperature of 65°C.
- Uses Bca-Optimized Taq™, an LA technology-based enzyme specially optimized (for use with BcaBEST™ DNA Polymerase.)
- The kit includes Random 9 mers or oligo-dT Primers for first strand synthesis. A specific downstream primer should be prepared by the user.
- The whole procedure is carried out in a single tube. The reverse transcription is stopped via heat inactivation of the enzyme and the PCR reaction mix is added directly to the tube and amplified.
Description:
The BcaBEST™ RNA PCR Kit is designed to perform both reverse transcription and DNA amplification in a single tube. The kit uses BcaBEST™ DNA Polymerase, which contains both DNA polymerase and reverse transcriptase for first strand cDNA synthesis. In contrast to standard reverse transcriptases, BcaBEST™ Polymerase has a temperature optimum of 65°C, which permits cDNA synthesis from difficult and highly structured RNA templates.
Bca-Optimized Taq™ Polymerase is used for second strand synthesis and subsequent PCR. This polymerase utilizes LA PCR technology for improved PCR length and accuracy. Random 9-mers, oligo-dT primers or a specific downstream primer that acts as an antisense primer in PCR can be used for cDNA synthesis.
Kit Components:
| TAK RR023A | |
| BcaBEST™ DNA Polymerase (22 U/μL) | 50 μL |
| RNase Inhibitor (40 U/μL) | 25 μL |
| Random 9-mers (50 μM) | 50 μL |
| Oligo dT Primer (50 μM) | 50 μL |
| RNase-Free dH2O (DEPC-treated) | 1 mL |
| Bca-Optimized Taq™ (5 U/μL) | 25 μL |
| 2X Bca 1st Strand Buffer | 2 x 1.25 mL |
| 5X Bca 2nd Strand Buffer | 800 μL |
| dNTP Mixture (10 mM each) | 50 μL |
| MgSO4 (25 mM) | 500 μL |
| Control F-1 Primer (20 μmol/μL) | 25 μL |
| Control R-1 Primer (20 μmol/μL) | 25 μL |
| Positive Control RNA (1 μg/μL) | 25 μL |
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