Prime-a-Gene® Labeling System Technical Bulletin
Literature # TB049
The Prime-a-Gene® Labeling System is based on the method developed by Feinberg and Vogelstein, in which a mixture of random hexadeoxyribonucleotides is used to prime DNA synthesis in vitro from any linear double-stranded DNA template. With this method it is possible to generate probes of extremely high specific activity (>1 x 109 cpm/µg), even using DNA fragments cut from agarose gels. Since the input DNA serves as a template and remains intact during the reaction, minimal amounts of DNA (25ng) can be labeled to a high specific activity. Typically, greater than 60% of the labeled deoxyribonucleotide can be incorporated into the Lambda Control DNA using the labeling reaction described here. Incorporation may vary from 40–80% with other samples, depending on the template and reaction conditions used. Using a template greater than 500bp, probes generated with the Prime-a-Gene® Labeling System are generally 250–300bp in length and are suitable for a variety of applications.
Summary of Changes
The following changes were made to the 2/15 revision of this document:
1. The suggested circular filters in Section 5.A were updated to replace a discontinued item on the materials list.
2. The document design was updated.
Printed in USA. Revised 2/15.