diagnosticsgenome.com Home Molecular Regents PCR PreMix 2.5X Syber Green PCR-PreMix For Real Time PCR2.5X syber green HOT START PCR-PREMIX for Real-time PCR is an optimized, ready to use PCR mixture of Hot start Taq DNA Polymerase, PCR buffer, Syber green, MgCl2 and dNTP's. This pre-mix contains all components for real-time PCR for syber green, except DNA template and primers. The mixture is suitable for amplification of most of the DNA templates in Real Time PCR with syber green chemistry. Tested for absence of endodeoxyribonucleases and exodeoxyrinucleases and is also tested for amplidication of a single gene copy.
SPECIFICATIONS : Supplied as 2.5X, in liquid form so as to ensure that you can add your primers & Template DNA volumes to desired levels.
APPLICATIONS : Suitable for complex genomic cDNA templates, low copy number targets, Quantitation of viral/bacterial loads by Real Time PCR etc.
STORAGE : Recommended storage is at-20°C.
Dilution PROTOCOL for PCR :
| Component | 50 µl reaction Volume | 25 µl reaction volume | ||
| Volume | Final Conc. | Volume | Final Conc. | |
| 2.5 X Syber Green HOT START PCR Premix | 20 µl | 1 X | 10 µl | 1 X |
| Forward primer | Variable | 0.1-1 µM | Variable | 0.1-1 µM |
| Reverse Primer | Variable | 0.1-1 µM | Variable | 0.1-1 µM |
| Template DNA | Variable | 10pg-1 µg | Variable | 10pg-1 µg |
| Sterile Deionized water | Up to 50 µl | ---- | Up to25 µl | ---- |
| Suggested Cycling protocol : | |||
| Initial Denaturation: | 95°C | 5 minutes | |
| Cycling | 95°C | 20 secs | |
| Annealing | 30 secs | 40 cycles | |
| 72°C extension | 30 secs | ||
| Final Extension | 72°C | 5 minutes | |
For low Copies PCR reactions can be extended to 45 Cycles.
Packing : The above PCR premix is available in convenient small packs as well as large packs for bulk users.
The pre-mix is supplied with an additional vial of DNase Free water. Pre-mixes with various concs of MgCl2 are available as mentioned below. Generally the 2.5mM Pre-mix is considered as universal, however for further optimization for your parameters you can try Premixes with other Concs of MgCl2.
| Cat. No. | PRODUCT | Pack Size | Pack Insert |
| 110171 | 2.5 X Syber Green HOT START PCR-Premix: 50 rxns of 50µl or 100 rxns of 25µl 1.5 mM final Mg at 1 X Conc | 1ml (50 rxns 50µl) | 110171P |
| 110172 | 2.5 X Syber Green HOT START PCR-Premix: 100 rxns of 50µl or 200 rxns of 25µl 1.5 mM final Mg at 1 X Conc | 1ml x 2 (100 rxns 50µl) | 110172P |
| 110173 | 2.5 X Syber Green HOT START PCR-Premix: 50 rxns of 50µl or 100 rxns of 25µl 2.5 mM final Mg at 1 X Conc | 1ml (50 rxns 50µl) | 110173P |
| 110174 | 2.5 X Syber Green HOT START PCR-Premix: 100 rxns of 50µl or 200 rxns of 25µl 2.5 mM final Mg at 1 X Conc | 1ml x 2 (100 rxns 50µl) | 110174P |
| 110175 | 2.5 X Syber Green HOT START PCR-Premix: 500 rxns of 50µl or 1000 rxns of 25µl 2.5 mM final Mg at 1 X Conc | 1ml x 10 (100 rxns 50µl) | 110175P |