End-Point PCR: AccuAmp PCR Master Mix
Cat. No. |
Size |
Price |
Quantity |
11009-50 |
50 x 50-µl reactions (1.25 ml) |
$69.00 |
|
11009-200 |
200 x 50-µl reactions (4 x 1.25 ml) |
$269.00 |
|
11009-01k |
1000 x 50-µl reactions (25 ml) |
$1119.00 |
Benefits
- High PCR specificity for use with numerous different targets and templates.
- Outstanding sensitivity with robust performance.
- Easy reaction setup at room temperature.
- Convenient 2x master mix.
Description
AccuAmp PCR Master Mix is a 2X concentrated, ready-to-use reaction cocktail that contains all components, except primers and template, for high specificity PCR amplification of genomic DNA or cDNA targets. For accurate PCR amplification, it is crucial to reduce/ avoid any nonspecific products and primer–dimers generated during amplification. AccuAmp PCR Master Mix promotes highly specific annealing of primers to the PCR template through our unique combination of proprietary buffer, stabilizers, and specially modified Taq DNA polymerase, which prevents amplification of nonspecific products.
Storage
AccuAmp PCR Master Mix is stable for 1 year when stored at –20°C in a constant-temperature freezer. After thawing, mix thoroughly before using.
Quick-Reference Protocol
1. Thaw AccuAmp PCR Master Mix (2X), template, and primers. Mix the individual solutions.
2. Prepare a reaction mix.
Component |
Volume/Reaction |
Final Concentration |
AccuAmp PCR Master Mix (2X) |
25 µl |
1X |
Forward Primer |
Variable |
300 nM |
Reverse Primer |
Variable |
300 nM |
Template DNA or cDNA |
Variable |
Up to 1 µg/reaction |
Nuclease-free Water |
Variable |
|
Total Reaction Volume |
50 µl |
|
3. Mix the reaction mix thoroughly. After sealing each reaction, centrifuge reaction tubes briefly to avoid bubbles.
4. When using a thermal cycler with a heated lid, do not use mineral oil. Otherwise, overlay with 100 µl mineral oil.
5. Program your thermal cycler.
Step |
Temperature &Time |
Notes |
PCR initial step |
95°C, 5 min |
|
3-step cycling (20-40 cycles) |
a. 95°C, 15 sec (Denaturation) |
Annealing temperature: ~5°C below Tm of primers; Extension: for PCR products longer than 1 kb, use an extension time of 1 min per kb DNA. |
Final extension |
72°C, 10 min |
|
6. Place the PCR tubes or plates in the thermal cycler, and start the cycling program.
7. After amplification, analyze the PCR products, or store at -20°C.