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Sequence
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==Sequencing Protocol== | ==Sequencing Protocol== | ||
'''Protocol:''' | '''Protocol:''' | ||
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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, use up to 8ul of DNA; if less than 8ul, add autoclaved deionized water. | + | Depending on plasmid size and concentration, use up to 8ul of DNA; if less than 8ul, add autoclaved deionized water for volume. |
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| - | ''DNA Type DNA Size DNA Volume(total) Primer Volume (total)'' | + | '''DNA Type/ DNA Size/ DNA Volume(total)/ Primer Volume (total)''' |
| - | Plasmid <6kb 8ul (~400ng) 4ul (8pmol) | + | Plasmid/ <6kb/ 8ul (~400ng)/ 4ul (8pmol) |
| - | Plasmid | + | Plasmid/ 6-10kb/ 8ul (~600ng)/ 4ul (8pmol) |
| - | Plasmid >10kb 8ul (~800ng) 4ul (8pmol) | + | Plasmid >10kb/ 8ul (~800ng)/ 4ul (8pmol) |
| - | Purified 200-500bp 8ul (3-10ng) 4ul (8pmol) | + | Purified PCR/ 200-500bp/ 8ul (3-10ng)/ 4ul (8pmol) |
| - | + | ||
| - | Purified .5-1kb 8ul (5-20ng) 4ul (8pmol) | + | Purified PCR/ .5-1kb/ 8ul (5-20ng)/ 4ul (8pmol) |
| - | + | ||
| - | Purified 1-2kb 8ul (10-40ng) 4ul (8pmol) | + | Purified PCR/ 1-2kb 8ul (10-40ng) 4ul (8pmol) |
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Latest revision as of 01:00, 30 October 2008
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)
