Use of solubilizable acrylamide disulfide gels for isolation of DNA fragments suitable for sequence analysis

✍ Scribed by J.Norman Hansen

• Publisher: Elsevier Science
• Year: 1981
• Tongue: English
• Weight: 507 KB
• Volume: 116
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(81)90337-7

✦ Synopsis

Acrylamide gels which arc crosslinked by disultide bridges are found to be an excellent electrophoretic medium for DNA restriction fragments over a wide size range. The gels are solubilised by mercaptoethanol and the fragments are rapidly and conveniently recovered on DEAE~cellulose columns. Fragment recovery appears nearly quantitative. The gel formulation described here requires polymerization at a temperature of about 40°C. Recovered fragments arc active in all systems tested. including subsequent restriction, kinasing, ligation, nick translation. and sequencing by the method of A. M. Maxam and W. Gilbert (1980. in Methods in Ervymology (Grossman.