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Time Cycles Initial denaturation 98 3 min 1 Denaturation 98 10 sec 15 (decrease by 1 C per cycle) Gradient Annealing 70-55C 20 sec Extension 72 30 sec/kb Denaturation 98C 10 sec 20 Annealing 55C 20 sec Extension 72C 30 sec/kb Final extension 72 5 min 1 4Characteristics of Different PCR Protocols: Protocol Type Speed Specificity PCR Yield Detection Rate Two-Step Fastest High Medium High Three-Step Medium Medium Highest Medium Gradient Annealing Slow High Medium High $15.00 - $3,665.00 2 HotStart Fast PCR Master Mix (With Dye) Product description 2 HotStart PCR Master Mix contains heat-stabilized Taq DNA Polymerase modified with antibodies, adds strong elongation factor and optimized buffer system, and has super high amplification efficiency

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Composites Science and Technology, 70 (1), 116122

One powerful and commonly used approach is to tag your protein of interest with Glutathione S-Transferase (GST) and perform a GST pull-down to see what binds to the protein of interest