Team:Stockholm/22 September 2010
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==Andreas== | ==Andreas== | ||
===Gel verification=== | ===Gel verification=== | ||
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::'''***Note to self***'''<br /> | ::'''***Note to self***'''<br /> | ||
::''It might be possible that pSB1A2.RBS⋅yCCS clones have been mixed up with pEX.N-TAT⋅SOD⋅His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes. | ::''It might be possible that pSB1A2.RBS⋅yCCS clones have been mixed up with pEX.N-TAT⋅SOD⋅His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes. | ||
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+ | == Mimmi == | ||
+ | |||
+ | === SOD / SOD.his / his.SOD / yCCS === | ||
+ | |||
+ | {| | ||
+ | | colspan="3" | ''colony PCR'' | ||
+ | | rowspan="8" width="100" | | ||
+ | ! samples | ||
+ | | rowspan="8" width="100" | | ||
+ | ! colspan="2" | Conditions | ||
+ | | rowspan="3" | | ||
+ | |- | ||
+ | | colspan="3" | | ||
+ | | pEX.SOD x3 | ||
+ | ! time | ||
+ | ! °C | ||
+ | |- | ||
+ | ! mix | ||
+ | | (µl) | ||
+ | | x13 | ||
+ | | pEX.SOD.his x3 | ||
+ | | 5m | ||
+ | | 95 | ||
+ | |- | ||
+ | | mastermix | ||
+ | | 24.5 | ||
+ | | rowspan="5" | | ||
+ | | pEX.his.SOD x3 | ||
+ | | 30s | ||
+ | | 95 | ||
+ | | ) | ||
+ | |- | ||
+ | | DNA | ||
+ | | 0.5 | ||
+ | | pEX.yCCS x3 | ||
+ | | 30s | ||
+ | | 55 | ||
+ | | > 30 cycles | ||
+ | |- | ||
+ | | align="right" | tot | ||
+ | | 25µl | ||
+ | | rowspan="3" | | ||
+ | | 1m20s | ||
+ | | 72 | ||
+ | | ) | ||
+ | |- | ||
+ | | colspan="2" rowspan="2" | | ||
+ | | 5m | ||
+ | | 72 | ||
+ | |- | ||
+ | | oo | ||
+ | | 25 | ||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | ==== Gel ==== | ||
+ | |||
+ | [[Image:2010-09-22_pEX.SOD_+_yCCS.jpg|200px|thumb|left|]] | ||
+ | {| | ||
+ | ! well | ||
+ | ! width="120" | sample | ||
+ | ! well | ||
+ | ! sample | ||
+ | |- | ||
+ | | 1 | ||
+ | | ladder | ||
+ | | 9 | ||
+ | | ladder | ||
+ | |- | ||
+ | | 2 | ||
+ | | pEX.SOD.his 1 | ||
+ | | 10 | ||
+ | | pEX.SOD 1 | ||
+ | |- | ||
+ | | 3 | ||
+ | | pEX.SOD.his 2 | ||
+ | | 11 | ||
+ | | pEX.SOD 2 | ||
+ | |- | ||
+ | | 4 | ||
+ | | pEX.SOD.his 3 | ||
+ | | 12 | ||
+ | | pEX.SOD 3 | ||
+ | |- | ||
+ | | 5 | ||
+ | | pEX.his.SOD 1 | ||
+ | | 13 | ||
+ | | pEX.yCCS 1 | ||
+ | |- | ||
+ | | 6 | ||
+ | | pEX.his.SOD 1 | ||
+ | | 14 | ||
+ | | pEX.yCCS 2 | ||
+ | |- | ||
+ | | 7 | ||
+ | | pEX.his.SOD 1 | ||
+ | | 15 | ||
+ | | pEX.yCCS 2 | ||
+ | |- | ||
+ | | 8 | ||
+ | | pEX.SOD.his | ||
+ | | 16 | ||
+ | | - | ||
+ | |} | ||
+ | |||
+ | {{Stockholm/Footer}} |
Latest revision as of 11:05, 26 October 2010
Contents |
Andreas
Gel verification
Of 21/9 colony PCR samples
1 % agarose, 120 V
Expected bands
- pSB1A2.RBS⋅yCCS (pA.RBSy): 1018 bp
- pEX.N-TAT⋅SOD⋅His (pEX.nTSH): 735 bp
- pSB1C3.N-LMWP⋅SOD⋅His (pC.nLSH): 857 bp
- pSB1K3.N-LMWP⋅SOD⋅His (pK.nLSH): 857 bp
- pSB1C3.SOD⋅His (PC): 815 bp
Results
- pSB1A2.RBS⋅yCCS: Seemingly correct-sized bands for clones 2, 3 and 4.
- pEX.N-TAT⋅SOD⋅His: Seemingly correct-sized bands for clones 3 & 4.
- pSB1C3.N-LMWP⋅SOD⋅His: Seemingly correct-sized bands for clones 1 & 4.
- pSB1K3.N-LMWP⋅SOD⋅His: Seemingly correct-sized band for clone 1.
ON cultures
- 5 ml LB + appr. antibiotic (100 Amp, 25 Cm or 50 Km); 37 °C, 225 rpm
- pSB1A2.RBS⋅yCCS: clones 3 and 4
- pEX.N-TAT⋅SOD⋅His: clones 3 and 4
- pSB1C3.N-LMWP⋅SOD⋅His: clones 1 and 4
- pSB1K3.N-LMWP⋅SOD⋅His: clone 1
- ***Note to self***
- It might be possible that pSB1A2.RBS⋅yCCS clones have been mixed up with pEX.N-TAT⋅SOD⋅His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes.
- ***Note to self***
Mimmi
SOD / SOD.his / his.SOD / yCCS
colony PCR | samples | Conditions | ||||||
---|---|---|---|---|---|---|---|---|
pEX.SOD x3 | time | °C | ||||||
mix | (µl) | x13 | pEX.SOD.his x3 | 5m | 95 | |||
mastermix | 24.5 | pEX.his.SOD x3 | 30s | 95 | ) | |||
DNA | 0.5 | pEX.yCCS x3 | 30s | 55 | > 30 cycles | |||
tot | 25µl | 1m20s | 72 | ) | ||||
5m | 72 | |||||||
oo | 25 |
Gel
well | sample | well | sample |
---|---|---|---|
1 | ladder | 9 | ladder |
2 | pEX.SOD.his 1 | 10 | pEX.SOD 1 |
3 | pEX.SOD.his 2 | 11 | pEX.SOD 2 |
4 | pEX.SOD.his 3 | 12 | pEX.SOD 3 |
5 | pEX.his.SOD 1 | 13 | pEX.yCCS 1 |
6 | pEX.his.SOD 1 | 14 | pEX.yCCS 2 |
7 | pEX.his.SOD 1 | 15 | pEX.yCCS 2 |
8 | pEX.SOD.his | 16 | - |
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