PCR-Sure Kit, 12 x 10 Reactions Download PDF Store at -20° C
| Product | Quantity | Cat. No |
| PCR-SureTM Kit | 12 x 5 rxns | G065 |
| Individual Reaction Mix | 200 rxns | G065-X* |
* There are a total of 12 Individual Reaction Mixes. After the optimal reaction condition is identified, the Individual Reaction Mixes may be ordered separately. For the catalog number G065-X, the “X” indicates a number from 1 to 12. When ordering, please specify a unique catalog number.
Product Description It has been well established that many variables affect a particular PCR reaction: template
structures, primer design, annealing temperature, concentration of Mg2+, etc. The PCR-Sure™
Kit was developed to simplify the PCR optimization process. The system consists of multiple
thermo-stable DNA polymerases pre-mixed with 12 optimized buffers in a 2X MasterMix
format, saving a great amount of time in PCR set-up compared to the non-MasterMix PCR
optimization format. To find the optimal conditions for your difficult PCR, all you need to do is
to mix templates, primers, and H2O with the PCR-Sure™ 2X MasterMix.
Quality Control Free of endonucleases, exonucleases, and nicking activity. Every lot is tested for performance
consistency.
Storage Store all components at -20°C in a non-frost-free freezer. All components are stable for 1 year from the date of shipping when stored and handled properly.
Experimental Procedures There are 12 different PCR reaction buffers and thus a reaction mix of 14 is generally needed.
| | 1X | 14X |
| Template (DNA) | 1 µl (50 - 100 ng) | 14 µl |
| Forward Primer (10 µ) | 1 µl | 14 µl |
| Reverse Primer (10 µ) | 1 µl | 14 µl |
| H2O | 23 µl | 322 µl |
Mix the components well and aliquot 25 μl to each of 12 PCR reaction tube, then add 25 μl
PCR-Sure™ 2X MasterMix to the 12 PCR tubes.
PCR Cycle Profile As is often the case, this kit is used only when you have a difficult PCR reaction, we recommend
that you use a touch-down PCR cycle profile to optimize your primer annealing temperature.
For example, if your primer annealing temperature is around 55oC, use the following PCR
cycle program. Adjust accordingly if your primer annealing temperature is higher or lower
than 55
oC.
Initial DNA denaturation: 94ºC for 4 minutes.
- Denature at 94° C for 30 secs
- Anneal at 60° C for 30 secs
- Extension at 72° C for 1 min/kb | 3 Cycles |
- Denature at 94° C for 30 secs
- Anneal at 55° C for 30 secs
- Extension at 72° C for 1 min/kb | 3 Cycles |
- Denature at 94° C for 30 secs
- Anneal at 50° C for 30 secs
- Extension at 72° C for 1 min/kb | 25 Cycles |
Final extension at 72° C for 5 minutes.
Equipment 