albumin - publications

Predict more albumin - ligand interactions now!

1. BMC Res Notes. 2012 May 24;5(1):257. [Epub ahead of print]

Bovine serum albumin further enhances the effects of organic solvents on
increased yield of polymerase chain reaction of GC-rich templates.

Farell EM, Alexandre G.

ABSTRACT: BACKGROUND: While being a standard powerful molecular biology
technique, applications of the PCR to the amplification of high GC-rich DNA
samples still present challenges which include limited yield and poor specificity
of the reaction. Organic solvents, including DMSO and formamide, have been often
employed as additives to increase the efficiency of amplification of high GC
content (GC > 60%) DNA sequences. Bovine serum albumin (BSA) has been used as an
additive in several applications, including restriction enzyme digestions as well
as in PCR amplification of templates from environmental samples that contain
potential inhibitors such as phenolic compounds. FINDINGS: Significant increase
in PCR amplification yields of GC-rich DNA targets ranging in sizes from 0.4 kb
to 7.1 kb were achieved by using BSA as a co-additive along with DMSO and
formamide. Notably, enhancing effects of BSA occurs in the initial PCR cycles
with BSA additions having no detrimental impact on PCR yield or specificity. When
a PCR was set up such that the cycling parameters paused after every ten cycles
to allow for supplementation of BSA, combining BSA and organic solvent produced
significantly higher yields relative to conditions using the solvent alone. The
co-enhancing effects of BSA in presence of organic solvents were also obtained in
other PCR applications, including site-directed mutagenesis and overlap extension
PCR. CONCLUSIONS: BSA significantly enhances PCR amplification yield when used in
combination with organic solvents, DMSO or formamide. BSA enhancing effects were
obtained in several PCR applications, with DNA templates of high GC content and
spanning a broad size range. When added to the reaction buffer, promoting effects
of BSA were seen in the first cycles of the PCR, regardless of the size of the
DNA to amplify. The strategy outlined here provides a cost-effective alternative
for increasing the efficiency of PCR amplification of GC-rich DNA targets over a
broad size range.

PMID: 22624992 [PubMed - as supplied by publisher]