Objective
- Re-quantify plasmid+insert minipreps from previous week
- BamH1 digest of above-mentioned samples.....
- Run gel with BamH1 digested samples
Bench work
- Analytical digest
- 7μL pTXB1 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1+BSA#1 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1+BSA#2 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1+BSA#3 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1His + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1His+BSA#1 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1His+BSA#1 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- 7μL pTXB1His+BSA#1 + 1μL 10X BSA + 1μL 10X NEB3 + 1μL BamHI
- Gel (with agarose at 0.8%)
- mix 2μL loading buffer with each sample from step 1 above and load on gel in same order, with 1KB ladder in the middle
- Double-checked miniprep [DNA] (User:Daniel Catt)
- Transformed ligation product from 7/7/11 into competent DH10B made on 7/15/11 using standard transformation protocol (User:Daniel Catt)
Results
Analytical gel
- All miniprep samples are the wrong size. This implies that the ligation failed entirely.
DNA concentrations
- New [DNA] on miniprep samples from 7/12/11
(Though DNA.....
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