The BcaBEST DNA Labeling Kit is a novel random primer labeling system that uses longer primers (9-mers) and BcaBEST polymerase, a thermostable DNA polymerase isolated from Bacillus caldotenax. Unlike E. coli Klenow fragment or modified T7 DNA polymerase, Bca polymerase allows stable polymerization of DNA even for templates with high GC content containing highly structured regions. Longer 9-mer primers increase the incorporation efficiency of labeled nucleotides up to 80%.
With this kit, random primer labeling via thermostable Bca polymerase is accomplished in 10 minutes, resulting in high yields of labeled DNA probe. The kit is designed for use with [32P]-alpha-, [35S]-alpha- or [3H]-alpha-dCTP labels, but it can also be used with nonradioactive biotin, digoxigenin, or DNP-labeled dUTPs through dNTP replacement. In general, probes with specific activities of >109 dpm/mg DNA obtained with [32P]-alpha-dCTP (~3,000 Ci/mmol, 111 TBq/mmol).