AmpliTaq Gold® 360 Master Mixby Applied Biosystems
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AmpliTaq Gold® 360 Master Mix for End-Point PCR – Applied Biosystems
Applied Biosystems’s AmpliTaq Gold® 360 Master Mix utilizes the enzyme AmpliTaq Gold 360 DNA Polymerase, to amplify Genomic DNA, cDNA, and similar templates. Included PCR reagents and buffers help support Hot-start PCR in addition to Multiplex PCR, allowing complete control over amplification specificity (i.e., specific or non-specific amplification) and complexity (single or multiple target amplification).
Polymerase and Protocol Specifications
The AmpliTaq Gold 360 DNA Polymerase is supplied at the specified concentration and is activated by conditions stated in the protocol during denaturing. Following annealing, nucleic acid samples of the suggested weight or volume are elongated at 72 °C. Applied Biosystems recommends a 50 µl PCR reaction volume and amplicons of < 5 kb for optimal polymerase performance. Utilizing thermal cycling and Hot-start PCR to replicate DNA, AmpliTaq Gold 360 DNA Polymerase extends primers with free dNTPs, forming a complementary second strand.
Applications & Downstream Experiments of AmpliTaq Gold® 360 Master Mix
Intended uses of Applied Biosystems’s AmpliTaq Gold® 360 Master Mix include PCR, Amplification of low-copy amplicons and long targets, as well as related PCR cloning and amplification techniques. The resulting product is applicable for use in sequencing, and other tested methods.
PCR Troubleshooting Tips
Contamination and inappropriate primer construction create amplification inefficiencies. Therefore, PCR cleanup kits enhance the purity of either the starting or final sample, maximizing accuracy, and effectiveness. Dedicated primer design tools may also be useful in those respects. Optimum temperatures suggested by Invitrogen should be enforced, e.g., elongation (72 °C). Fidelity and enzyme concentration (see product specifications) also need to be accounted for when setting up your PCR.