Thermostable DNA polymerases

Thermostable DNA polymerases

• GSure™  Ultra Nucleic Acid Kit for DNA & RNA Isolation
• GSure™ Dogma kit for DNA, RNA & Protein Isolation
• Reverse Transcription & cDNA Synthesis
• Thermostable DNA Polymerases
• Fastract™Direct PCR Kit
• SYBR green & Taqman based qRT-PCR Kits
• TA-Cloning kit and Related Enzymes
• Competent Cells
• Site Directed Mutagenesis Based Kits
• Sequencing Grade Reagents

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^xeroPolymerase

Description

'xero' in Greek means dry or with little water. ^xeroPolymerase from GCC Biotech is a dried 2X ready load Taq master mix. This is the first time in global market, a ready load 2X master mix is coming as a dried form. To set the reaction, one need to recover the master mix with nuclease free water (provided with the ^xeroPolymerase separately) and add template and primers. The master mix is 'ready to load', so no need to add loading dye before running the PCR products in gel, making the master mix more convenient and decreasing the chances of loading error. After electrophoresis two tracking dyes would be visible. Orange colour dye migrating parallel to 50bp double stranded DNA and blue coloured dye migrating with 4 Kb double stranded DNA(in 0.7-1% agarose gel).

The mastermix is compatible for amplification upto 3 Kb (Figure 1) efficiently. This was also checked that for higher size amplification, efficiency increases with longer (1.5min/kb)  ncubation at extension step within the cycle (data not shown).

^xeroPolymerase master mix contains such amount of Taq DNA polymerase that change in reaction volume would hardly make any difference in amplification efficiency (Figure 2).

The ^xeroPolymerase are available in different formats. For ready to go pack, 8 tubes strip and 96 well plates are available. Each tube or well contains ^xeroPolymerase for the reaction volume of 20ml. Another pack of the ^xeroPolymerase is also available where 50 reactions, each of 20ml reaction volume are lyophilized in the screw capped tube. For 96 well plate format or 8 tubes strip format, before using the ^xeroPolymerase, only nucleases free water, template and primers are required to be added in the tube. Any further mixing by vortexing is not at all required, tap mixing by finger is well enough for efficient amplification (Figure 3).

Storage stability of the ^xeroPolymerase has been checked at 4^oC, room temperature (25^oC -30^oC) and at 37^oC. This has been proved that the ^xeroPolymerase is stable at 4^oC at least for 6 months while storage at room temperature is recommended for not more than 15- 20 days. Storage at room temperature decreases the polymerase efficiency for longer amplicons after 15 days (Figure 4A and 4B). Increasing the extension time in the cycle overcomes this efficiency fall (data not shown). ^xeroPolymerase, at 37^oC is only stable for first five days. Further storage at 37^oC drastically reduces the efficiency of amplification for longer amplicons, up to 1kb, severity observed after 15 days of storage (Figure 4^oC).

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Protocol & Manuals

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GX1001P	XeroPolymerase Dry PCR Mix, (1 plate X 96 well Plate), 96 rxn		GX1001P	XeroPolymerase Dry PCR Mix, (1 plate X 96 well Plate)96 rxn
GX1002P	XeroPolymerase Dry PCR Mix, (5 plate X 96 well Plate), 5 X 96 rxn		GX1002P	XeroPolymerase Dry PCR Mix, (5 plate X 96 well Plate), 5 X 96 rxn
GX1001S	XeroPolymerase Dry PCR Mix, (8 tube strip with cap) (10 strip), 80 rxn		GX1001S	XeroPolymerase Dry PCR Mix, (8 tube strip with cap) (10 strip), 80 rxn
GX1002S	XeroPolymerase Dry PCR Mix, (8 tube strip with cap) (100 strip), 80 rxn		GX1002S	XeroPolymerase Dry PCR Mix, (8 tube strip with cap) (100 strip), 80 rxn
GX1001T	XeroPolymerase Dry PCR Mix Tube, 50 rxn		GX1001T	XeroPolymerase Dry PCR Mix Tube, 50 rxn
GX1002T	XeroPolymerase Dry PCR Mix Tube, 4 X 50 rxn		GX1002T	XeroPolymerase Dry PCR Mix Tube, 4 X 50 rxn
GX1003T	XeroPolymerase Dry PCR Mix Tube, 12 X 50 rxn		GX1003T	XeroPolymerase Dry PCR Mix Tube, 12 X 50 rxn