Promega
GoTaq® Green Master Mix
M7123
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I routinely use CRISPR/Cas9-based methods to generate gene knockouts in cell lines. Following CRISPR/Cas9-mediated editing with a dual-guide approach, I derive single-cell clonal lines and identify edited clones by PCR using Promega's GoTaq PCR Master Mix.
Molecular Biology
Genotyping sample prepared using QuickExtract DNA Extraction Solution
None
1. PCR mixture (20 µL): 10 µL of 2× GoTaq Green Master Mix, 0.4 µL each of forward and reverse primer (10 µM stock; 0.2 µM final), 1 µL of template (from a 50 µL lysate prepared from a confluent 96-well clonal line), and nuclease-free water to 20 µL. 2. Cycling conditions: initial denaturation at 95 °C for 2 min; 35 cycles of 95 °C for 30 s, 60 °C for 30 s, and 72 °C for 30 s; final extension at 72 °C for 5 min; hold at 12 °C. 3. A 10 µL aliquot of each reaction was resolved on a 1.5% agarose gel in TAE buffer.
Ease of use and product performance
Highly recommended for genotyping PCRs