IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/27: Difference between revisions
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==Team Flavor== | ==Team Flavor== | ||
===Ligation of Mira/Brazz + StrepII to STOP + V0120=== | |||
*Insert to Backbone ration of 3:1 | |||
Miraculin: 34ng | |||
Brazzein: 11ng | |||
Backbone: 50ng | |||
*Ligations were transformed, plated onto LB+AMP and left overnight @ 37°C | |||
===PCR of Valencene using Pfx Polymerase=== | |||
*Pfx polymerase was rumoured to be more accurate at PCR from genomic DNA | |||
Specs: | |||
DNA 1-1: 1.8 ng/μL | |||
DNA 1-2: 1.1 ng/μL | |||
DNA 2-1: 2.6 ng/μL | |||
DNA 2-2: 2.2 ng/μL | |||
*1μL of each DNA sample was used for PCR. Cycling procedure can be found here: http://www.invitrogen.com/etc/medialib/en/filelibrary/pdf.Par.79411.File.dat/platinumpfx_pps.pdf | |||
===Mira/Brazz+StrepII+STOP confirmation digest=== | |||
Digested DNA: | |||
MN2: 211.2 ng | |||
MC2: 190.2 ng | |||
BN2: 184.4 ng | |||
BC2: 219.2 ng | |||
*Digestions were run on a 1% agarose gel at 125V for 30 minutes. | |||
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Revision as of 15:20, 27 July 2010
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Team Allergy
1-6: LTP + PDK (3200 & 1200); 7-16: LTP + PAL (3200 & 540); 17,18: Bet1 + PDK (3200 & 1200); 19-22: Bet1 + PAL (3200 & 540); 23,24: Bet2+PDK (3200 & 1200); 25-28: Bet2 + PAL (3200 & 540); 29-32: Ger3 + PDK (3200 & 1200); 33-38: Ger3 + PAL (3200 & 540); Image: Ladder (every 6)
Antisense-Intron Backbone Digestion
(Let it go for 2 hours and then phosphatized backbone (20 min)
Team FlavorLigation of Mira/Brazz + StrepII to STOP + V0120
Miraculin: 34ng Brazzein: 11ng Backbone: 50ng
PCR of Valencene using Pfx Polymerase
Specs: DNA 1-1: 1.8 ng/μL DNA 1-2: 1.1 ng/μL DNA 2-1: 2.6 ng/μL DNA 2-2: 2.2 ng/μL
Mira/Brazz+StrepII+STOP confirmation digestDigested DNA: MN2: 211.2 ng MC2: 190.2 ng BN2: 184.4 ng BC2: 219.2 ng
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