High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences

High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences.

Isolation of unknown DNA sequences flanked by known sequences is an important task in molecular biology research. Thermal asymmetric interlaced PCR (TAIL-PCR) is an effective method for this purpose. However the success rate of the original TAIL-PCR needs to be increased, and it is more desirable to obtain target products with larger sizes. Here we present a substantially improved TAIL-PCR proc...

Single-primer amplification of flanking sequences.

“megaprimer method” of site-directed mutagenesis. BioTechniques 8:404-407. 11.Senanayake, S.D. and D.A. Brian. 1995. Precise large deletions by the PCR based overlap extention. Mol. Biotechnol. 4:13-15. 12.Silver, J., T. Limjoco, and Feinstone. 1995. Site-specific mutagenesis using the polymerase chain reaction, p. 179-188. In M.A. Innis, D.H. Gelfand and J.J. Sninsky (Eds.), PCR Strategies. Ac...

Thermal asymmetric interlaced PCR amplification of YAC insert end fragments for chromosome walking in Plasmodium falciparum and other A/T-rich genomes.

Recombinant DNA sequences generated by PCR amplification.

The polymerase chain reaction (PCR) has greatly enhanced the field of molecular biology by making numerous regions of the genome (coding and noncoding), in both extant and extinct taxa, accessible for detailed analysis. PCR is especially well suited for applications in systematic biology because conserved regions that flank variable portions of the genome can be used as primer sites for the amp...

Improved PCR method for amplification of GC-rich DNA sequences.

Most housekeeping genes, tumor-suppressor genes, and approx 40% of tissue-specific genes contain G+C sequences in their promoter region that were very difficult to amplify. In this report, we propose an improved polymerase chain reaction (PCR) method to be used for successful amplification of the tissue factor pathway inhibitor (TFPI)-2 gene promoter region that exhibit >70% G+C content in a se...