AmpliTaq Gold™ DNA Polymerase with Buffer II and MgCl2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.

Publication protocol

PCR amplification
DNA samples were extracted using QIAamp DNA Blood Mini Kit (Qiagen, Valencia, CA) and the concentration was determined using NanoDrop ND-100 spectrophotometer (NanoDrop Technologies, Wilmington, DE). PCR reactions were performed in a 10 μl total volume containing 2 pmol of forward primer, 2 pmol reverse primer, 2.5 mM MgCl2, 0.2 mM each deoxyribonucleotide, 0.25 units AmpliTaq Gold DNA polymerase and 1.0 μl buffer (Applied Biosystems, Foster City, CA). Samples containing 10 ng DNA were subjected to 35 cycles of denaturation (95°C, 30 sec), annealing (52°C, 30 sec) and extension (72°C, 60 sec). The total amount of forward primers and reverse primers used for multiplex PCR was both 5 pmol and the AmpliTaq Gold DNA polymerase was increased to 0.5 units.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for AmpliTaq Gold™ DNA Polymerase with Buffer II and MgCl2 below.

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