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Team:Freiburg/Notebook/12 July
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=Sequence Analysis= | =Sequence Analysis= | ||
==<span style="color:orange;">Lysis cassette</span>== | ==<span style="color:orange;">Lysis cassette</span>== | ||
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==<span style="color:green;">green light receptor</span>== | ==<span style="color:green;">green light receptor</span>== | ||
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| - | + | Comments: Miniprep from June,12 were loaded onto a gel. | |
Ccas (M31, M32, M30)in pSB1C3 | Ccas (M31, M32, M30)in pSB1C3 | ||
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Image of the gel of M30-MM32 and M36-M38 | Image of the gel of M30-MM32 and M36-M38 | ||
| - | + | Comments: All the samples were negative. | |
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| - | + | Miniprep from June,12 were loaded onto a gel. If they are positive: Sequencing | |
ho1+terminator (M36, M37, M38) in pSB1C3 | ho1+terminator (M36, M37, M38) in pSB1C3 | ||
| - | + | Comments: All the samples were negative. See image above. | |
==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
Latest revision as of 00:55, 22 September 2011
Contact 
Contents |
Sequence Analysis
Lysis cassette
Lysis cassette and heat inducible promotor
S15b: (Lysis cassette K124014) is named K124017 in the registry Part without RBS at the beginning Part without antiholin the sequece is also mutated leading to wrong amino acids if combined with RBS there would be too many bases (8) between RBS and coding sequence part is likely to be not effective
green light receptor
CcaR
S17: (CcaR with Prefix and RBS) is ok. Check RBS. S18: seems to be ok too.
red light receptor
pycA+terminator
S24: prefix and suffix not complete
cph8
S29: Sequence without part. Has to be done again.
Digestion
| Name: | Date: |
| Continue from Date Name
Experiment | |
| Project Name: | |
Procedure
- add H2O (38μl-DNA )
- 5 μl NEB4 buffer (stored at iGEM’s, -20°C)
- 5 μl 10x BSA (used 1:10 diluted sample stored at iGEM’s, -20°C)
- DNA (500 ng) (Vector:Insert ratio 1:3 in following Ligation)
- 1 μl restriction enzymes (stored at iGEM’s, -20°C)
- heat for 1-2 hours 37°C (6 hours if time)
- heat for 20 minutes 80°C (inactivation of enzymes)
- keep at 4°C if you cannot continue
Measured DNA-concentration with Nanodrop to calculate the volume of DNA to do the digestion:
| Sample Name | DNA concentration (μg/μl) |
| P20 GFP | 132 |
| P19 GFP | 145 |
| P18 GFP | 107 |
| S39 PR | 90 |
| S43 PR | 40 |
| Tet Vector | 25 |
blue light receptor
LOV1
S33: LOVtap only
LOV3
S35: is K322999. This sequence is the LOVtap with the tryptophane promoter, RBS, mRFP and two terminators.
green light receptor
Gel
Investigators: Jakob
Comments: Miniprep from June,12 were loaded onto a gel.
Ccas (M31, M32, M30)in pSB1C3
Image of the gel of M30-MM32 and M36-M38
Comments: All the samples were negative.
red light receptor
Gel
Investigators: Jakob
Miniprep from June,12 were loaded onto a gel. If they are positive: Sequencing
ho1+terminator (M36, M37, M38) in pSB1C3
Comments: All the samples were negative. See image above.
Precipitator
Plastic binding site
Investigators: Sophie
Picking colonies (M2a, M2b, M3), inoculating LB (Cm) and let grow overnight. Start: 17:15
