Biotechrabbit은 바이오의약품 및 생명과학 연구를 위한 고급 실험 재료와 서비스를 제공하는 회사입니다.
제품 설명
2X YourTaq™ PCR Master Mix
제품 번호
BR0102201 - 200 reactions of 50 µl
BR0102202 - 1000 reactions of 50 µl
BR0102203 - 4000 reactions of 50 µl
제품 특징
Description
biotechrabbit™ YourTaq PCR Master Mix is optimized for high yield of amplification of 0.1–3 kb DNA targets, even from low copy number. The mix shows excellent PCR specificity and sensitivity for a broad range of amplicons. YourTaq PCR Master Mix is resistant to PCR inhibitors, such as blood (up to 20%), Ethanol or humic acid enabling PCR amplification from DNA templates with carry-over of PCR-inhibitors.
The 2X YourTaq PCR Master Mix contains pure biotechrabbit YourTaq DNA Polymerase, extremely high-quality dNTPs and optimized PCR buffer; thus, only template, PCR primers and PCR-grade water are added. In addition, the mix is suitable for amplification of GC-rich templates (up to 70%) pairing with 5X PCR Enhancer (BR1900201).
Info: Recommended annealing temperature is 2°C above primer Tm (use gradient PCR to optimize the annealing temperature).
Other Information
Quality ControlFunctional assayHuman genomic DNA was amplified using the PCR Master Mix and specific primers to produce a distinct band. |
Protocols
When assembling the amplification reactions, care should be taken to eliminate the possibility of contamination with undesired DNA.
The standard PCR protocol using biotechrabbit reaction buffer provides excellent results for most applications. Optimization might be necessary for certain conditions, such as the amplification of long targets, high GC or AT content, strong template secondary structures or insufficient template purity. In such cases, optimization of template purification (see biotechrabbit nucleic acid purification kits), primer design and annealing temperature is recommended.
The best conditions for each primer-template can be optimized with the following:
Component | Volume | Final concentration |
2X YourTaq PCR Master Mix | 25 µl | 1× |
5X PCR Enhancer (optional - BR1900201) | 10 µl | 1× |
Forward primer | Variable | 0.2–1 µM |
Reverse primer | Variable | 0.2–1 µM |
Template DNA | Variable | 10 pg–1 μg |
| Use 0.01–1 ng for plasmid or phage DNA and 0.05–1 μg for genomic DNA | |
Nuclease free water | Variable |
|
Total volume | 50 µl |
|
Step | Temperature | Time | Cycles |
Initial activation | 95°C | 2 min | 1 |
Denaturation | 95°C | 30 s | 25–35 |
Annealing* | (55-68°C) | 15–30 s | 25–35 |
| *Recommended annealing temperature is 2°C above Tm of primers. | ||
Extension | 72°C | 30–60 s/kb | 25–35 |
Final extension | 72°C | 5 min | 1 |
| To extend all incomplete PCR products | ||
Storage in the cycler | 4°C | Indefinitely | 1 |
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