ROMAX Plus, 2X which is a highly enhanced version of ROMAX 2X provides a complete ingredient needed for PCR include; Taq DNA polymerase, PCR buffer, dNTPs, except DNA template and primers. By reducing pipetting steps, ROMAX Plus, 2X help to reduce contamination. The enzyme consists of a single polypeptide with molecular weight of approximately 94 k Da. It has 5’-3’ DNA polymerase activity and 5’-3’ exonuclease activity. It lacks 3’-5’ exonuclease activity. This enzyme is suitable for routine amplification.
- High throughput PCR.
- Routine PCR
- Generation of PCR products for TA. Cloning
- Thawing ROMAX Plus, 2X; then gently invert and briefly centrifuge.
- Place PCR tube on ice and add following component to each tube.
|ROMAX Plus, 2X||25 µl|
|Forward primer (10 µM)||1 µl|
|Reverse primer (10 µM)||1 µl|
|Water for Molecular Biology||Up to 50 µl|
|Total volume||50 µl|
- Spin down the PCR tube and run the PCR program.
|Initial denaturation||95 ̊C||1-3 min||1|
|Denaturation||95 ̊C||30 s|| |
Certificate of analysis
- Endodeoxyribonuclease Assay
No detectable degradation of DNA was observed after incubation of 1 µg of pUC19 DNA with 25 µL of ROMAX Plus, 2X in 50 µL of reaction mixture for 4 hours at 37°C and at 70°C.
- Exodeoxyribonuclease Assay No detectable degradation of DNA was observed after incubation of 1 μg of lambda DNA/HindIII fragments with 25 μL of ROMAX Plus (2X) in 50 μL of reaction mixture for 4 hours at 37°C and at 70°C.
- Ribonuclease Assay
No contaminating RNase activity was detected after incubation of 1 µg of [3H]-RNA with 25 µL ROMAX Plus, 2X in 50 µL of reaction mixture for 4 hours at 37°C and at 70°C.
- Functional Assay
ROMAX Plus, 2X was tested for amplification of 1,110 bp single copy gene from human genomic DNA.
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