IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/14: Difference between revisions
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===DTL of StrepII+Mira/Brazz with STOP codon=== | ===DTL of StrepII+Mira/Brazz with STOP codon=== | ||
*To insert a STOP codon to the end of our Mira/Brazz StrepII constructs, we used the STOP+V0120 BioBrick as our vector (EcoR1/Xba1 digest) and our Miraculin/Brazzein construct as our insert (EcoR1/Spe1 digest) | |||
*Construct DNA used was from Miniprep 1 (Box 4) | |||
{| border="1" | |||
|+ ''Digestion Reactions'' | |||
! !! Mira Strep N !! Mira Strep C !! Brazz Strep N !! Brazz Strep C !! STOP+V0120 BioBrick | |||
|- | |||
! diH<sub>2</sub>O | |||
| align="center" | 15 || align="center" | 14.5 || align="center" | 13.5 || align="center" | 11.5 || align="center" | 6 | |||
|- | |||
! Green FD buffer (10x) | |||
| align="center" | 2 || align="center" | 2 || align="center" | 2 || align="center" | 2 || align="center" | 2 | |||
|- | |||
! DNA | |||
| align="center" | 1 || align="center" | 1.5 || align="center" | 2.5 || align="center" | 4.5 || align="center" | 10 | |||
|- | |||
! EcoR1 | |||
| align="center" | 1 || align="center" | 1 || align="center" | 1 || align="center" | 1 || align="center" | 1 | |||
|- | |||
! Xba1 | |||
| align="center" | - || align="center" | - || align="center" | - || align="center" | - || align="center" | 1 | |||
|- | |||
! Spe1 | |||
| align="center" | 1 || align="center" | 1 || align="center" | 1 || align="center" | 1 || align="center" | - | |||
|- | |||
|} | |||
''Digestion of Mira/Brazz+Strep and STOP codon BioBrick'' <br/> | ''Digestion of Mira/Brazz+Strep and STOP codon BioBrick'' <br/> |
Revision as of 12:52, 14 July 2010
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Team FlavorMiniprep of STOP + V0120 partNanodrop O/D's: STOP V0120 1-1: 134.4 STOP V0120 1-2: 98.7 STOP V0120 2-1: 55.2 STOP V0120 2-2: 121.8 DTL of StrepII+Mira/Brazz with STOP codon
Digestion of Mira/Brazz+Strep and STOP codon BioBrick
Team FencePCR of the arabidopsis promoters with the newly extracted DNA and the older DNA. FAIL Arp2 promoter PCR failed. Exp2 promoter PCR failed. Phenol Chloroform DNA Extraction
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