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Sequence
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Revision as of 00:30, 29 October 2008
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 PCR/ 200-500bp/ 8ul (3-10ng)/ 4ul (8pmol)/
Purified PCR/ .5-1kb/ 8ul (5-20ng)/ 4ul (8pmol)/
Purified PCR/ 1-2kb 8ul (10-40ng) 4ul (8pmol)
