diagnosticsgenome.com Home Molecular Regents Single Tube X-AMP Universal 2.5X Single Tube RT/PCR PreMixX-AMP Universal 2.5 X Single tube RT/PCR PREMIX is an optimized, stabilized ready to use RT/PCR mixture of RT enzyme, Hot Start Taq DNA Polymerase, RT/PCR buffer, MgCl2 and dNTP's besides some additives for enhances amplifications except RNA template and primers. The 2.5x RT/PCR-premix contains all components for cDNA synthesis and subsequent PCR in a single tube. The mixture is suitable for amplification of most of the RNA templates. Tested for absence of endodeoxyribonucleases and exodeoxyribonucleases.
SPECIFICATIONS : Supplied as 2.5X in liquid form so as to ensure that you can add your primers and Template RNA volumes to desired levels.
APPLICATIONS : Suitable for complex RNA templates, low copy number targets, large number of thermal Cycles, multiplex PCR. Quantitation of viral loads by Real Time PCR etc.
STORAGE : Recommended storage is at - 20°C.
SUGGESTED DILUTION PROTOCOL FOR SINGLE TUBE RT/PCR :
| Component | 50 µl reaction Volume | 25 µl reaction volume | ||
| Volume | Final Conc. | Volume | Final Conc. | |
| 2.5 x 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 cDNA synthesis the target specific primers can be used.
Packing : The above RT/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 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 |
| 110181 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 50 rxns of 50µl or 100 rxns of 25µl 1.5 mM final Mg at 1 X Conc | 1ml (50 rxns 50µl) | 110181P |
| 110182 | X-AMP Universal SINGLE TUBE 2.5 X RT/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) | 110182P |
| 110183 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 50 rxns of 50µl or 100 rxns of 25µl 2.5 mM final Mg at 1 X Conc | 1ml (50 rxns 50µl) | 110183P |
| 110184 | X-AMP Universal SINGLE TUBE 2.5 X RT/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) | 110184P |
| 110185 | X-AMP Universal SINGLE TUBE 2.5 X RT/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) | 110185P |
| 110186 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 50 rxns of 50µl or 100 rxns of 25µl 3.5 mM final Mg at 1 X Conc | 1ml (50 rxns 50µl) | 110186P |
| 110187 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 100 rxns of 50µl or 200 rxns of 25µl 3.5 mM final Mg at 1 X Conc | 1ml x 2 (100 rxns 50µl) | 110187P |
| 110188 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 50 rxns of 50µl or 100 rxns of 25µl 4 mM final Mg at 1 X Conc | 1ml (50 rxns 50µl) | 110188P |
| 110189 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 100 rxns of 50µl or 200 rxns of 25µl 4 mM final Mg at 1 X Conc | 1ml x 2 (100 rxns 50µl) | 110189P |
| 110190 | X-AMP Universal SINGLE TUBE 2.5 X RT/PCR-Premix: 100 rxns of 50µl or 200 rxns of 25µl 5 mM final Mg at 1 X Conc | 1ml x 2 (100 rxns 50µl) | 110190P |
| 110191 | 2.5 X RT/PCR-Premix: 100 rxns of 50µl or 200 rxns of 25µl with special additives for specialized applications | 1ml x 2 (100 rxns 50µl) | 110191P |