"
Sequence
From 2008.igem.org
(New page: ==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...) |
(→Sequencing Protocol) |
||
| Line 6: | Line 6: | ||
Sequencing primers should be diluted to 5uM; 4.0ul of 5uM primer should be used. | Sequencing primers should be diluted to 5uM; 4.0ul of 5uM primer should be used. | ||
| - | Depending on plasmid size and concentration | + | 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 | ||
Revision as of 00:27, 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 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
