12/09/13
From 2013.igem.org
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|'''DNA volume'''||'''Volume'''||'''Cutsmart buffer'''||'''Enzyme volume'''||'''dH2O added to total volume of 60 ul''' | |'''DNA volume'''||'''Volume'''||'''Cutsmart buffer'''||'''Enzyme volume'''||'''dH2O added to total volume of 60 ul''' | ||
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- | |TODX||23|6||1||30 | + | |TODX||23||6||1||30 |
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|TODF||22.5||6||1||30.5 | |TODF||22.5||6||1||30.5 |
Revision as of 20:26, 13 September 2013
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Contents |
Isolate plasmid from overnight culture
- Using Omega mini prep kit protocol
- Samples:
- TODF 1,2,3,
- TODX 1, 2, 3
- TOBG 1, 2, 3,
- RFP 1, 2, 3
sample | Volume (ul) | Concentration (ng/ul) |
TODF1 | 43 | 120 |
TODF2 | 43 | 98.8 |
TODX1 | 42 | 399.3 |
TODX2 | 44 | 442.2 |
TODX3 | 43 | 417.5 |
TOBG1 | 43 | 262.4 |
TOBG2 | 47 | 319.2 |
TOBG3 | 41 | 338.4 |
RFP1 | 41 | 77.7 |
RFP2 | 41 | 32.7 |
RFP3 | 42 | 59 |
Glycerol stocks
- Took 750ul from overnight culture and centrifuged at full speed for ~2 minutes
- Supernatant was removed
- Pellet resuspended in 375ul of HMFM
- Samples were then frozen at -80
Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13
The ge was purified by using the The Zymoclean Gel DNA recovery Kit and its protocol was followed.
Nanodrop Results
Sample | Volume | Concentration (ng/ul) | 280/260 | 260/230 |
TODX | 48.5 | 7.1 | 1.83 | 0.72 |
TODF | 45.5 | 21.7 | 1.89 | 1.19 |
pSB1C3 (RFP) | 46.2 | 7.3 | 1.81 | 0.16 |
Digestion of the samples purified with the enzyme PstI-HF
DNA volume | Volume | Cutsmart buffer | Enzyme volume | dH2O added to total volume of 60 ul |
TODX | 23 | 6 | 1 | 30 |
TODF | 22.5 | 6 | 1 | 30.5 |
pSB1C3 (RFP) | 20.5 | 6 | 1 | 32.5 |