Publication Cover
Hemoglobin
international journal for hemoglobin research
Volume 12, 1988 - Issue 5-6
9
Views
1
CrossRef citations to date
0
Altmetric
Original Article

Restriction Primer Extension Method of Labeling Oligonucleotide Probes and Its Application to the Detection of HB E Genes

, , , , , , & show all
Pages 691-697 | Received 22 Dec 1987, Accepted 01 Jun 1988, Published online: 07 Jul 2009
 

Abstract

A new method for labeling oligonucleotides was developed to obtain high specific activity of radioactive probes. In an oligonucleotide molecule, two sequences were designed. One sequence, the 5′, contains 19 nucleotides and serves as a template for probe synthesis. The second sequence, 3′, contains a consensus sequence which forms a Pst I site after forming a complementary strand with the primer. In the presence of E. coli DNA polymerase I (Klenow fragment), α-32P dNTP and other dNTPs, a radioactive labeled oligonucleotide was synthesized by the primer extension method. After Pst I digestion, the probe was different from its template in length by 4 bp and could be separated from each other on urea-polyacrylamide gel electrophoresis (PAGE). A radioactive 01ionucleotide probe with extremely high specific activity up to 1010 dpm/μg could be obtained by the use of this method. The oligonucleotide probes have been used for the detection of the Hb E mutation in this report.

Reprints and Corporate Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

To request a reprint or corporate permissions for this article, please click on the relevant link below:

Academic Permissions

Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?

Obtain permissions instantly via Rightslink by clicking on the button below:

If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.