User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/07/20

AU CHEM-570 Lab Prep

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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 Digest miniprepped plasmids with BamHI to confirm size (it cuts once, so size should be ~8.5kb) 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 1h 20' @ 37°C 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.....