Adding carrier nucleic acids to extraction buffers increases the yield of extracted DNA and RNA from analytical samples. Poly-A used as a carrier molecule enables and enhances many molecular biology methods, protocols, and tools, for example, by stabilizing extracted pathogenic RNA or synthetic RNA or DNA, for instance, large oligonucleotide molecules.
According to Shaw et al., the addition of a poly-A carrier RNA to chaotropic salt solutions increases oligonucleotide yields. The researchers observed that the use of nucleic acid carrier molecules enhances DNA extraction methods in microfluidic applications.
Poly A carrier molecules are also quite useful for the stabilization of other RNA molecules during gene editing studies. For example, Miyaoka et al. employed a poly-A carrier when quantifying repair of double-strand breaks during the homology-directed repair. Miyaoka et al. developed a rapid, digital PCR-based assay that can simultaneously detect one HDR or NHEJ event out of 1,000 copies of the genome. This assay allowed monitoring genome-editing outcomes of CRISPR-associated protein 9 (Cas9), Cas9 nickases, catalytically dead Cas9 fused to FokI, and transcription activator-like effector nuclease at three disease-associated endogenous gene loci in HEK293T cells, HeLa cells, and human-induced pluripotent stem cells.
Jacobsen N, Nielsen PS, Jeffares DC, Eriksen J, Ohlsson H, Arctander P, Kauppinen S. Direct isolation of poly(A)+ RNA from 4 M guanidine thiocyanate-lysed cell extracts using locked nucleic acid-oligo(T) capture. Nucleic Acids Res. 2004 Apr 19;32(7):e64. [PMC]
Miyaoka Y, Berman JR, Cooper SB, et al. Systematic quantification of HDR and NHEJ reveals effects of locus, nuclease, and cell type on genome-editing. Sci Rep. 2016;6:23549. [PMC]
Shaw KJ, Thain L, Docker PT, et al. The use of carrier RNA to enhance DNA extraction from microfluidic-based silica monoliths. Anal Chim Acta. 2009;652(1-2):231-233. [Article]
Bio-Synthesis Inc. is pleased to offer a large variety of DNA/RNA oligonucleotides and peptides, modified or unmodified including poly A and poly T oligonucleotides, various conjugates, as well as stapled peptides and regular peptides, for a number of research applications, including COVID 19 testing, analysis and vaccine development!