Sequence

Revision as of 00:27, 29 October 2008 by Lphalen (Talk | contribs)

Sequencing Protocol

Protocol:

Go to Genewiz.com to order sequencing.

Sequencing primers should be diluted to 5uM; 4.0ul of 5uM primer should be used.

Depending on plasmid size and concentration, use up to 8ul of DNA; if less than 8ul, add autoclaved deionized water.

Specifications According to Genewiz.com:

DNA Type DNA Size DNA Volume(total) Primer Volume (total)

Plasmid <6kb 8ul (~400ng) 4ul (8pmol)

Plasmid 6-10kb 8ul (~600ng) 4ul (8pmol)

Plasmid >10kb 8ul (~800ng) 4ul (8pmol)

Purified 200-500bp 8ul (3-10ng) 4ul (8pmol) PCR

Purified .5-1kb 8ul (5-20ng) 4ul (8pmol) PCR

Purified 1-2kb 8ul (10-40ng) 4ul (8pmol) PCR PCR