IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/26: Difference between revisions
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We digested the vectors with Spe and Pst1 and ligate with antisense parts (already digested and gel purified). We ran them on the E Gel too long so the lanes do not show anything conclusive, other than that there is digested DNA in the bottom half. | We digested the vectors with Spe and Pst1 and ligate with antisense parts (already digested and gel purified). We ran them on the E Gel too long so the lanes do not show anything conclusive, other than that there is digested DNA in the bottom half. | ||
Result: [[Image:2-assembly_digest.jpg|right|200px]] | Result: [[Image:2-assembly_digest.jpg|right|200px]] | ||
'''Sequence review''' clarified which parts of the sequences on the team page corresponded to sense/antisense. | '''Sequence review''' clarified which parts of the sequences on the team page corresponded to sense/antisense. | ||
Revision as of 08:20, 30 July 2010
 iGEM iGarden
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Team FencePFX polymerase PCR
9.5μL total DNA: 21ng/μL
9.5μL + 4μL = 13.5μL therefore 6.5μL H20 per reaction MM
total volume:100μL Arabidopsis PCR
Team AllergyProceduresLast week, we finished ligation together Sense and Intron parts of the hpRNA and transformed them into E. coli. This week, we will extract the plasmids, add in the antisense part, and transform. Procedures today...
We also need to check which parts of the sequence on the team page corresponds to the sense/antisense parts of the gene. There may have been mistakes in the past. ResultsCulture We cultured 38 colonies. All grew but one LTP+PAL and Ger+Pal grew. We labeled the cultures 1- 38 for simplicity sake. 1-6: LTP + PDk 7-16: Miniprep Here are the concentrations of the miniprepped plasmid.
Diagnostic Digest We digested the vectors with Spe and Pst1 and ligate with antisense parts (already digested and gel purified). We ran them on the E Gel too long so the lanes do not show anything conclusive, other than that there is digested DNA in the bottom half. Result: Sequence review clarified which parts of the sequences on the team page corresponded to sense/antisense. Team Flavor
Digestion Reactions
Mira N: 888ng
Mira C: 888ng
Brazz N: 993ng
Brazz C: 1032ng
STOP: 781ng x 2 (max DNA available)
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