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제품 설명
2X YourTaq™ Direct-Load Hot-Start PCR Master Mix
제품 번호
BR0202301 - 200 reactions of 50 µl
BR0202302 - 1000 reactions of 50 µl
BR0202303 - 4000 reactions of 50 µl
제품 특징
Description
biotechrabbit™ YourTaq Direct-Load Hot Start PCR Mix is optimized for high yield of amplification of 0.1–3 kb DNA targets, even from low copy number. YourTaq Hot Start PCR Mix shows excellent PCR specificity and sensitivity for a broad range of amplicons. The 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 2× YourTaq Direct-Load Hot Start PCR Mix contains pure biotechrabbit YourTaq Hot Start DNA Polymerase, extremely high-quality dNTPs, two dyes (blue and yellow) that separate during electrophoresis, allowing migration progress to be monitored, and sufficient buffer density for direct loading onto agarose gels. In addition, the mix is suitable for amplification of GC-rich templates (up to 70%) pairing with 5× PCR Enhancer (BR1900201).
Info: Recommended annealing temperature is 2°C above primer Tm (use gradient PCR to optimize the annealing temperature).
Other Information
Component | Composition |
YourTaq Direct-Load Hot Start PCR Mix | Optimized 2× YourTaq Hot Start PCR Master Mix containing electrophoresis tracking dyes (yellow and blue) and density reagent for direct gel loading |
STORAGE | −20°C (until expiry date – see product label) |
Human 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 |
YourTaq Direct-Load Hot Start PCR Mix, 2× | 25 µl | 1× |
5× 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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