Application of blocking oligonucleotides to improve signal-to-noise ratio in a PCR.

Hege Vestheim, Bruce E. Deagle, Simon N. Jarman

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)

Abstract

"Universal" or group-specific PCR primers have a tendency to predominately hybridise with the common sequences in samples with mixed templates. The result is that the rarer sequences are seldom retrieved by cloning or sequencing. The use of a blocking oligonucleotide (oligo) designed to specifically prevent amplification of dominant or unwanted DNA templates is an easy way to improve the amplification of rarer sequences. Here, we describe the different types of blocking principles and the different types of blocking oligos and give guidelines and examples of their application.

Original languageEnglish
Pages (from-to)265-274
Number of pages10
JournalMethods in molecular biology (Clifton, N.J.)
Volume687
Publication statusPublished - 1 Jan 2011

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