Excellent and fun learning exercise on long product high GC PCR. We’ve gone the gene synthesis route these days - would’ve costed $300-400 including sequence verification and initial cloning into a plasmid vector. The learning experience here is priceless. Well done!
Come with me as I troubleshoot this very high GC PCR toward clonable homology arms in an actinobacteria. Testing DMSO, Betaine 1M, and Propylene Glycol to see if we get amplification. Arms are 1000bp. Full length with arms and gene is 2800bp. Let's go!
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