Sequence

From 2008.igem.org

(Difference between revisions)
(Sequencing Protocol)
(Sequencing Protocol)
Line 12: Line 12:
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''DNA Type    DNA Size    DNA Volume(total)    Primer Volume (total)''
+
'DNA Type/     DNA Size/     DNA Volume(total)/     Primer Volume (total)'
   
   
    
    
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Plasmid      <6kb        8ul (~400ng)          4ul (8pmol)
+
Plasmid/     <6kb/         8ul (~400ng)/         4ul (8pmol)/
-
Plasmid     6-10kb       8ul (~600ng)          4ul (8pmol)
+
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)/    
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PCR
+
-
Purified    .5-1kb      8ul (5-20ng)          4ul (8pmol)
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Purified PCR/     .5-1kb/       8ul (5-20ng)/         4ul (8pmol)/
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PCR
+
-
Purified    1-2kb        8ul (10-40ng)        4ul (8pmol)
+
Purified PCR/     1-2kb        8ul (10-40ng)        4ul (8pmol)
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PCR     
+
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PCR
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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)