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DTI CobaltAmp HiFid Taq HS Premix
DTI CobaltAmp HiFid Taq HS Premix is a Fast PCR Master Mix and designed for streamlined PCR, and is, therefore, ideal for applications such as colony PCR. It contains a hot-start PCR enzyme, optimized buffer, dNTPs, along with a blue gel-loading dye and density reagent, in a convenient 2X premix format.
Reactions are assembled by simply mixing primers and DNA template with DTI CobaltAmp HiFid Taq HS Premix. For rapid and convenient screening, the amplification reactions can be loaded directly onto an agarose gel immediately after amplification without purification, or digested directly with restriction enzymes.
Brand | Catalogue Number | Pack Size |
DTI CobaltAmp HiFid Taq HS Premix | DT0203.80 | 80 rxn (25 μl volume) |
DTI CobaltAmp HiFid Taq HS Premix | DT0203.320 | 320 rxn (25 μl volume) |
• Colony PCR can be completed in half the time required for conventional Taq DNA polymerase.
• Convenient protocol eliminates purification steps with no reduction in yield.
• Restriction enzyme digestion of the PCR products can be performed directly in the PCR buffer.
• Reactions can be directly loaded onto a gel without purification or the addition of other reagents.
Results
Speed up Colony PCR with DTI CobaltAmp HiFid Taq HS Premix PCR Master Mix
Amplicon length | DTI CobaltAmp HiFid Taq HS Premix PCR Master Mix | Company P dye-added master mix | Company T dye-added master mix |
0.5 kb | 50 min | 1 hr 35 min | 1 hr 30 min |
1.0 kb | 55 min | 1 hr 50 min | 1 hr 45 min |
2.0 kb | 1 hr | 2 hr 20 min | 2 hr 15 min |
4.0 kb | 1 hr 30 min | 3 hr 20 min | 3 hr 15 min |
Experimental example: Using DTI CobaltAmp HiFid Taq HS Premix PCR Master Mix to screen a cDNA library by colony PCR
E. coli cells were transformed with a mouse cDNA library (insert size from 0.5 to 5 kb) ligated into pUC118. Plasmid-containing transformants were analyzed using DTI CobaltAmp HiFid Taq HS Premix PCR Master Mix and M4 and RV primers (see Methods below). The PCR reaction cycle was complete in just 70 minutes, and excellent yields were obtained. All 16 clones checked were confirmed to contain 0.5- to 5-kb inserts.
Figure 2. Colony PCR with DTI CobaltAmp HiFid Taq HS Premix PCR Master Mix.
Methods
DTI JadeAmp FabTaq Premix can amplify targets up to 10 kb in length, including targets that are GC- or AT-rich. Additionally, PCR products generated with DTI JadeAmp FabTaq Premix can be used directly for restriction enzyme digestion, sequencing, or TA-cloning without need for further purification.
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