Team:Michigan/Virus Surface Display
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- | 7/10/2010 | + | '''7/10/2010''' |
13:00 | 13:00 | ||
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- | 7/13/2010 | + | '''7/13/2010''' |
9:00 | 9:00 | ||
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- | 7/15/2010 | + | '''7/15/2010''' |
4:30 | 4:30 | ||
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- | 7/18/2010 | + | '''7/18/2010''' |
5:00 | 5:00 | ||
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- | 7/21/2010 | + | '''7/21/2010''' |
6:00 | 6:00 | ||
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- | 7/23/2010 | + | '''7/23/2010''' |
9:00 | 9:00 | ||
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- | 7/25/2010 | + | '''7/25/2010''' |
5:00 | 5:00 | ||
• aliquot INP samples and primers for sequencing | • aliquot INP samples and primers for sequencing | ||
- | 7/26/2010 | + | |
+ | '''7/26/2010''' | ||
+ | |||
• Electroporation transformation of the following Biobrick parts: | • Electroporation transformation of the following Biobrick parts: | ||
- | pBad/araC (I0500) | + | 1. pBad/araC (I0500) |
- | Linker (K157013) | + | 2. Linker (K157013) |
- | GFP (E0040) | + | 3. GFP (E0040) |
+ | |||
- | 7/27/2010 | + | '''7/27/2010''' |
9:00 | 9:00 | ||
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- | 7/28/2010 | + | '''7/28/2010''' |
4:00 | 4:00 | ||
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[[Image:7-28biobrickgel.jpg|300px]] | [[Image:7-28biobrickgel.jpg|300px]] | ||
- | Lane 1 (far right): | + | Lane 1 (far right): Invitrogen 1 kb Plus ladder |
Lane 2: GFP cut with XbaI and PstI | Lane 2: GFP cut with XbaI and PstI | ||
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- | 7/30/2010 | + | '''7/30/2010''' |
10:00 | 10:00 | ||
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• Heat shock transformation of ligation | • Heat shock transformation of ligation | ||
- | 8/2/2010 | + | |
+ | '''8/2/2010''' | ||
• Miniprep of INP-linker | • Miniprep of INP-linker | ||
• Nanodrop results: | • Nanodrop results: | ||
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2.1 59.8 1.99 2.06 | 2.1 59.8 1.99 2.06 | ||
2.2 69.8 2.02 2.11 | 2.2 69.8 2.02 2.11 | ||
+ | |||
+ | '''8/3/2010''' | ||
+ | |||
+ | • EcoRI+SpeI and SpeI+PstI digests of INP + linker minipreps | ||
+ | |||
+ | • Gel electrophoresis results: | ||
+ | |||
+ | [[Image:080310gel.jpg|250px]] | ||
+ | |||
+ | Lane 1 - 1 kb Plus Ladder | ||
+ | |||
+ | Lane 2 - INP + Linker 1.1 (EcoRI+SpeI) | ||
+ | |||
+ | Lane 3 - INP + Linker 1.2 (EcoRI+SpeI) | ||
+ | |||
+ | Lane 4 - INP + Linker 2.1 (EcoRI+SpeI) | ||
+ | |||
+ | Lane 5 - INP + Linker 2.2 (EcoRI+SpeI) | ||
+ | |||
+ | Lane 6 - INP + Linker 1.1 (SpeI+PstI) | ||
+ | |||
+ | Lane 7 - INP + Linker 1.2 (SpeI+PstI) | ||
+ | |||
+ | Lane 8 - INP + Linker 2.1 (SpeI+PstI) | ||
+ | |||
+ | Lane 9 - INP + Linker 2.2 (SpeI+PstI) | ||
+ | |||
+ | Lane 10 - INP 1 (EcoRI+SpeI) | ||
+ | |||
+ | Lane 11 - INP 2 (SpeI+PstI) | ||
+ | |||
+ | |||
+ | '''8/5/2010''' | ||
+ | |||
+ | • Inoculated 6 ml DH5α for ligation and transformation. | ||
+ | |||
+ | |||
+ | '''8/6/2010''' | ||
+ | |||
+ | • OmpA and GFP Ligations done: | ||
+ | 1. OmpA1 (insert) + GFP2 (backbone) | ||
+ | 2. OmpA2 (backbone) + GFP1 (insert) | ||
+ | |||
+ | • Heat shock transformation | ||
+ | |||
+ | |||
+ | '''8/9/2010''' | ||
+ | |||
+ | • Miniprep of 8/6 transformation, in addition to pBAD promoter and linker. | ||
+ | • Nanodrop results: | ||
+ | |||
+ | Sample OD260/280 OD260/230 ng/uL | ||
+ | 1. pBAD 1.72 8.91 11.5 | ||
+ | 2. OmpA+GFP 1 2.88 -3.52 4.5 | ||
+ | 3. OmpA+GFP 2 1.62 3.03 7.9 | ||
+ | 4. Linker 2.10 -12.00 9.4 | ||
+ | |||
+ | • 20 uL digest without water | ||
+ | - 5 uL NEB2 buffer | ||
+ | - 0.5 uL 100X BSA | ||
+ | - 1 uL of each enzyme | ||
+ | - 12.5 uL of DNA | ||
+ | |||
+ | • Digests: | ||
+ | 1. Linker 1 (EcoRI+SpeI) | ||
+ | 2. Linker 2 (EcoRI+SpeI) | ||
+ | 3. OmpA+GFP 1 (XbaI+PstI) | ||
+ | 4. OmpA+GFP 2 (XbaI+PstI) | ||
+ | 5. pBAD (SpeI+PstI) | ||
+ | |||
+ | |||
+ | '''8/10/10''' | ||
+ | |||
+ | • Gel of 8/9 digest results | ||
+ | |||
+ | [[Image:081010gel.jpg|350px]] | ||
+ | |||
+ | |||
+ | '''8/12/2010''' | ||
+ | |||
+ | • Due to poor digest results, we repeated the miniprep from 8/9, with additional colonies of OmpA+GFP picked. Also miniprepped are 2 expression plasmids (Backbone and Promoter) transformed earlier by another group. | ||
+ | |||
+ | Sample | ||
+ | |||
+ | |||
+ | |||
|width="250px" style="background: transparent"| | |width="250px" style="background: transparent"| | ||
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[[Image:Virus05.jpg|middle|250px]] | [[Image:Virus05.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus06.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus07.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus08.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus09.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus10.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus11.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus12.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus13.jpg|middle|250px]] | ||
+ | |||
+ | [[Image:Virus14.jpg|middle|250px]] | ||
+ | |||
|} | |} |
Latest revision as of 04:44, 27 October 2010