Technique / Molecular Biology / RNA transcriptional and post-transcriptional regulation / Primer extension

Primer extension is the fastest way to determine the transcriptional initiation site.

Primer extension assay for RNA analysis protocol
This is a PDF protocol from Marimoto lab at Northwestern University. References: Dignam et al. 1983. Nuc. Acids Res. 11, 1475-1489. Dynan and Tijian. 1985 Nature 316, 774-778. ...

Improved efficiency for primer extension by using a long, highly-labeled primer protocol
A open access technical article. Nucleic Acids Research, Vol 25, Issue 8 1658-1659 Improved efficiency for primer extension by using a long, highly-labeled primer generated from immobilized single-stranded DNA templates G Flouriot, C Pope, MR Kenealy ...

Primer Extension Protocol protocol
Primer extension protocol for highly structured RNAs (Genomics facility, University of Edinburgh). - Oligonucleotide purification - Oligo labeling - Primer extension ...

In vitro transcription and primer extension protocol
The following reaction conditions can be thought of as "standard" conditions based on procedures from the Kadonaga and Biggin labs. The nuclear extracts are prepared from Drosophila embryos according to Mark Biggin's protocol, which yield protein concentratio ...

Primer extension analysis of RNA 5' ends protocol
Primer extension method from Jim Brown's hypercard stack. The sites of transcription initiation or RNA processing are generally determined by S1 protection or primer extension (PE) analysis. Although S1 protection must be used for 3'-end analysis, 5'-end an ...

Primer-Extension Preamplified DNA Is a Reliable Template for Genotyping review
A open access technical article on genotyping based on primer extension amplifed DNA template. Clinical Chemistry 48: 1601-1603, 2002 Primer-Extension Preamplified DNA Is a Reliable Template for Genotyping Helena Kuivaniemi1,2, Sungpil Yoon1, Hidenori ...


More primer extension protocols

• References
• Related Books
• Discussion
• Related news
• Job search

Primer extension.
(Curr Protoc Mol Biol. 2001 May;Chapter 4:Unit4.8.)
Triezenberg SJ.
Michigan State University, East Lansing, Michigan, USA.

This protocol can be used to map the 5' terminus of an RNA and to quantitate the amount of a given RNA by extending a primer using reverse transcriptase. The primer is an oligonucleotide (or restriction fragment) that is complementary to a portion of the RNA of interest. The primer is end-labeled, hybridized to the RNA, and extended by reverse transcriptase using unlabeled deoxynucleotides to form a single-stranded DNA complementary to the template RNA. The resultant DNA is analyzed on a sequencing gel. The length of the extended primer maps the position of the 5' end of the RNA, and the yield of primer extension product reflects the abundance of the RNA.