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Team:TorontoMaRSDiscovery/11 June 2009
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June 11, 2009
- We did not have enough isolated plasmid DNA to justify doing another gel so we decided to perform another gel so we decided to perform another miniprep to get some more
- Used DB3.1 transformed bacteria from shaker(measured spec 0.794)
- Miniprep
- potential issue our microcentrifuge only spins @ 10,000 not required 12,000
- Binding DNA
- did both optional steps preheated TE + washed w/ W10
- measured UV absorbance = 4.2ng/ul
- in all there is ~ 74 ul of TE should indicated ~370 ng of plasmid.
- Digest
- NOTE: plasmid length w/e biobric is 3189 bp biobrick length is 3255bp this can explain lack of 2 clear bands on gel after ligation.
- nevertheless, we will perform another Gel to make sure we can properly run ladder to determine fragment sizes
- digest measurements: 250ng plasmid all else half except BSA
