Amplification of GC-rich DNA for high-throughput family-based genetic studies

摘要: Researchers face a significant problem in PCR amplification of DNA fragments with high GC contents. Analysis these regions is importance since many regulatory different genes and their first exons are GC-rich. There large number protocols for GC-rich DNA, some which perform well but costly. Most the economical fail to consistently, especially on products >80 % contents size >300 bp. One requires multiple additions polymerase during thermal cycling therefore rules out its utility if samples have be amplified. We established method simultaneous specific from human using general laboratory reagents. These amplicons ranging 65–85 sizes up 870 The protocol uses buffer containing co-solvents including 2-mercaptoethanol bovine serum albumin DNA. A profile also used incorporates annealing temperature 7 cycles reactions. suitable molecular biology applications sequencing.