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Team:Cornell/Week 7
From 2011.igem.org
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==Wednesday== | ==Wednesday== | ||
| + | |||
| + | Set up ligation reaction of pZE-12 backbone with the primer dimer insert. | ||
| + | Digestion of Backbone | ||
| + | - 26 ul Backbone plasmid = 2 ug DNA | ||
| + | - 5 ul Buffer #4 (optimal for SphI-HF and ClaI-HF) | ||
| + | - 1 ul SphI & 1 ul ClaI | ||
| + | - 16.5 ul H20 | ||
| + | left in water bath for one hour | ||
| + | - 1 ul CIAP added to digestion reaction | ||
| + | left in water bath for 30 minutes | ||
| + | |||
| + | Ran Digestion Product through gel for 30 minutes at 120 V. | ||
| + | Lane 1: 2 ul 1kb Ladder + 2 ul 6X Dye + 8 ul H20 | ||
| + | Lane 2: 25 ul digestion reaction + 5 ul 6X Dye | ||
| + | Lane 3: 25 ul digestion reaction + 5 ul 6X Dye | ||
| + | |||
| + | Gel Purification using Qiagen Kit | ||
| + | |||
| + | NanoDrop of Samples | ||
| + | Concentration: | ||
| + | Label: | ||
| + | |||
| + | Ligation Reaction | ||
| + | Label: | ||
| + | |||
| + | Ligation done overnight for transformation on 7/21/11 | ||
==Thursday== | ==Thursday== | ||
Revision as of 19:32, 20 July 2011
Week 1 | Week 2 - 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 | Week 15 | Week 16 | Week 17 | Week 18 | Week 19 | Week 20 | Week 21 |
July 17th - July 23rd
Sunday
- Successfully miniprepped 1 sample of GFP + avitag (the second sample got messed up)
- Ran a gel on the RFP that was PCRed on Saturday. We also cut out the bands.
- Sean will innoculate 6 tubes of GFP-containing bacteria--1 tube for us, and 5 tubes for CURIE.
Monday
- Gel purified 2 RFP PCR samples from Friday.
- Miniprepped 2 samples of GFP + avitag.
Tuesday
Submitted GFP+avitag genes for synthesis
Submitted pZE-12 backbone for subcloning of light sensor
Wednesday
Set up ligation reaction of pZE-12 backbone with the primer dimer insert.
Digestion of Backbone
- 26 ul Backbone plasmid = 2 ug DNA
- 5 ul Buffer #4 (optimal for SphI-HF and ClaI-HF)
- 1 ul SphI & 1 ul ClaI
- 16.5 ul H20
left in water bath for one hour
- 1 ul CIAP added to digestion reaction
left in water bath for 30 minutes
Ran Digestion Product through gel for 30 minutes at 120 V.
Lane 1: 2 ul 1kb Ladder + 2 ul 6X Dye + 8 ul H20
Lane 2: 25 ul digestion reaction + 5 ul 6X Dye
Lane 3: 25 ul digestion reaction + 5 ul 6X Dye
Gel Purification using Qiagen Kit
NanoDrop of Samples
Concentration:
Label:
Ligation Reaction
Label:
Ligation done overnight for transformation on 7/21/11
