Denaturing gel electrophoresis for sequencing.

摘要: The accuracy of DNA sequence determination depends largely upon resolution the sequencing products in denaturing polyacrylamide gels. This unit provides a detailed description setup, electrophoresis, and processing such In general, gels required for are 40-cm long, uniform thickness, contain 4% to 8% acrylamide 7 M urea. Modifications this protocol increase length readable information which can be obtained from single gel (i.e., forming with wedge-shaped spacers create field gradient, or incorporating buffer an electrolyte step gradient into gel). A modification Basic Protocol--inclusion formamide gel--is designed overcome compressions arising secondary structure during electrophoresis. discussion concentrations electrophoresis conditions is included Commentary.