IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/17: Difference between revisions
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#STOP codon + NOSterminator Sequence - 76.5 ng/μL | #STOP codon + NOSterminator Sequence - 76.5 ng/μL | ||
To do: follow up with restriction digest <font color="green"> ✓ </font> and Agarose gel to confirm PCR and PCR purification. <!--<font color="green"> ✓ </font>--> | To do: follow up with restriction digest <font color="green"> ✓ </font> and Agarose gel to confirm PCR and PCR purification. <!--<font color="green"> ✓ </font>--> | ||
===Restriction Digest=== | |||
Restriction Digest reactions were set up as follows: | |||
pENTCUP2 Promoter: | |||
27μL PCR product | |||
3μL Loading Buffer w/ dye | |||
1μL Xba1 Fast Enzyme | |||
1μL Pst1 Fast Enzyme | |||
NOSterm and NOSterm + STOP: | |||
9μL PCR product | |||
1μL Loading Buffer w/ dye | |||
0.5μL Xba1 Fast Enzyme | |||
0.5μL Pst1 Fast Enzyme | |||
BioBrick Plasmid V0120: | |||
3μL DNA | |||
1μL Loading Buffer w/ dye | |||
6μL diH<sub>2</sub>O | |||
0.5μL Xba1 Fast Enzyme | |||
0.5μL Pst1 Fast Enzyme | |||
Revision as of 11:03, 17 June 2010
 iGEM Project name 1
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Team FlavourPCR Purification of pORE Vector PartsFollowing QIAgen PCR Purification Kit Protocol the following PCR products were purified:
To do: follow up with restriction digest ✓ and Agarose gel to confirm PCR and PCR purification. Restriction DigestRestriction Digest reactions were set up as follows: pENTCUP2 Promoter: 27μL PCR product 3μL Loading Buffer w/ dye 1μL Xba1 Fast Enzyme 1μL Pst1 Fast Enzyme NOSterm and NOSterm + STOP: 9μL PCR product 1μL Loading Buffer w/ dye 0.5μL Xba1 Fast Enzyme 0.5μL Pst1 Fast Enzyme BioBrick Plasmid V0120: 3μL DNA 1μL Loading Buffer w/ dye 6μL diH2O 0.5μL Xba1 Fast Enzyme 0.5μL Pst1 Fast Enzyme
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