Melbourne/Lab Notebook Weeks 5-8
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-All lanes had a single band at 1000-1100bp. Two had a 1000bp band and three a 1100bp band. | -All lanes had a single band at 1000-1100bp. Two had a 1000bp band and three a 1100bp band. | ||
| - | *Colony 3 of above (1100bp band) was | + | *Colony 3 of above (1100bp band) was [[Melbourne/Miniprep protocol|Miniprepped]] using the Wizerd kit. |
*OD of miniprep measured using a [[Melbourne/DNA concentration measurement|spectrophotometer]] to be 20ng/ul. | *OD of miniprep measured using a [[Melbourne/DNA concentration measurement|spectrophotometer]] to be 20ng/ul. | ||
Revision as of 13:24, 8 October 2007
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Contents |
Week 5
23 July 2007
- Colony PCRed using vf2 and vR primers the transformations from 20/7. 3 colonies from ligation plate and one from plasmid control plate.
24 July 2007
- Ran Gel from PCRs (23/7) at 100V.
-results: colony 3 didn't give product.
- Streaked out some cells from index plate (No.3).
25 July 2007
26 July 2007
27 July 2007
Week 6
30 July 2007
(no BSA or AP used).
- Inactivated enzymes at 80degreesC in a heat block for 10min.
- Ligated the two parts together (5ul each) overnight at 4degreesC.
31 July 2007
- Transformation of ligations from 31/7 into NM522 competent cells using 3ul of DMSO.
using miniprep from 13/7. All digests were E/P.
-Results:
- Low digestion efficiency
- Low digestion efficiency
- No insert seen but DNA present
- No insert seen but DNA present
- Ok digestion (not very good)
- Ok digestion (not very good)
--Digestion imcomplete??
1 Aug 2007
- Cultured 6 colonies picked from 1/8 transformations. A total of about 500 colonies grew with none on control (competent cells only plate).
- Transformation of P2 9E (LacZa with KanR) from iGEM plate.
2 Aug 2007
- Alkaline Lysis Miniprepped colonies from 1/8. (P1 11A + P1 15P ligation. Named OMP-YFP)
-results: colony 6 had two bands of sizes approximately 1000 and 1100bp.
- Streaked out colony 6 liquid culture on LBA plate and grew overnight.
3 Aug 2007
- Picked 5 colonies from streaked out plate from 2/8 and Cultured overnight.
4 Aug 2007
- Alkaline Lysis Miniprepped colonies from 3/8.
-All lanes had a single band at 1000-1100bp. Two had a 1000bp band and three a 1100bp band.
- Colony 3 of above (1100bp band) was Miniprepped using the Wizerd kit.
- OD of miniprep measured using a spectrophotometer to be 20ng/ul.
Week 7
5 Aug 2007
6 Aug 2007
7 Aug 2007
8 Aug 2007
9 Aug 2007
10 Aug 2007
11 Aug 2007
Week 8
12 Aug 2007
13 Aug 2007
14 Aug 2007
15 Aug 2007
16 Aug 2007
17 Aug 2007
18 Aug 2007
