PCR with detachable primers

PCR with arbitrary primers

Sequencing-independent method to generate oligonucleotide probes targeting a variable region in bacterial 16S rRNA by PCR with detachable primers.

Oligonucleotide probes targeting the small-subunit rRNA are commonly used to detect and quantify bacteria in natural environments. We developed a PCR-based approach that allows synthesis of oligonucleotide probes targeting a variable region in the 16S rRNA without prior knowledge of the target sequence. Analysis of all 16S rRNA gene sequences in the Ribosomal Database Project database revealed ...

Designing PCR Primers Painlessly†

Improved PCR specificity with Hot Start PCR primers.

The polymerase chain reaction (PCR) is an indispensable DNA amplification technique that has been exploited in numerous areas, including molecular diagnostics. One major drawback of PCR is the competing amplification of undesired off-target products, which primarily occurs at ambient temperatures prior to PCR cycling (1). Hot Start PCR techniques aim to block the extension of primers in nonspec...

Selecting specific PCR primers with MFEprimer.

Selecting specific primers is crucial for polymerase chain reaction (PCR). Nonspecific primers will bind to unintended genes and result in nonspecific amplicons. MFEprimer is a program for checking the specificity of PCR primers against the background DNA. In this chapter, we introduce: (1) the factors that affect the specificity of primers; (2) the principle of MFEprimer and its settings; (3) ...