Sequence

From 2008.igem.org

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PRINCETON IGEM 2008

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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 for volume.


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)