NYMU Taipei/Lab Notes/2007 10 2
From 2007.igem.org
< NYMU Taipei/Lab Notes(Difference between revisions)
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** lane 18: OmpRBS (PCR 6) | ** lane 18: OmpRBS (PCR 6) | ||
== re-culture pOmpC and pCinRHSL == | == re-culture pOmpC and pCinRHSL == | ||
| - | == | + | * Because of the low concentration of pOmpC and pCinRHSL, the liquid culture is performed again to gain larger concentration |
| + | * concentrations | ||
| + | ** pCinRHSL (0.05 ug/uL by pett, 0.05 and 0.08 by 海威) | ||
| + | ** pOmpC #1 (run out off in previous experiment) | ||
| + | ** pOmpC #2 (0.02 ug/uL by pett, 0.04 and 0.05 by 海威) | ||
| + | ** pOmpC #3 (0.04 and 0.23 by 海威) | ||
| + | ** pOmpC #4 (0.04 and 0.08 by 海威) | ||
| + | |||
| + | == digestion for assembly == | ||
| + | === System 1 === | ||
| + | * pOmpC + TATA_INSA | ||
| + | ** pOmpC ('''re-cultured, 10/2'''; '''S.P. digestion, 10/3''') | ||
| + | ** TATA_INSA ('''X.P. digestion, 10/2''') | ||
| + | * <s>TATB_INSB + CinR+HSL+D-term | ||
| + | ** TATB_INSB ('''<font color=red>E.S. digestion, 10/2 (it is not correct!!! TATB_INSB does not have EcoRI site</font>''') | ||
| + | ** CinR+HSL+D-term ('''E. digestion, 10/2''') | ||
| + | ** CinR+HSL+D-term ('''X. digestion, 10/3''') | ||
| + | * (pOmpC + TATA_INSA) + (TATB_INSB + CinR+HSL+D-term) | ||
| + | ** (pOmpC + TATA_INSA) | ||
| + | ** (TATB_INSB + CinR+HSL+D-term)</s> | ||
| + | * pOmpC + TATA_INSA + TATB_INSB | ||
| + | * pOmpC + TATA_INSA + TATB_INSB + CinR+HSL+D-term | ||
| + | |||
| + | === System 2 === | ||
| + | * pCinRHSL + OmpRBS | ||
| + | ** pCinRHSL ('''re-cultured, 10/2'''; '''S.P. digestion, 10/3''') | ||
| + | ** OmpRBS ('''X.P. digestion, 10/2''') | ||
| + | * (pCinRHSL + OmpRBS) + TATD_IDE | ||
| + | ** (pCinRHSL + OmpRBS) | ||
| + | ** TATD_IDE | ||
| + | * [(pCinRHSL + OmpRBS) + TATD_IDE] + D-term | ||
| + | ** [(pCinRHSL + OmpRBS) + TATD_IDE] | ||
| + | ** D-term | ||
Latest revision as of 04:43, 9 October 2007
Contents |
TATA_INSA, TATB_INSB, TATD_IDE and OmpRBS GEL check after PCR
- from left to right
- lane 1: 100bp ladder
- lane 2-4: TATA_INSA (PCR 1+2)
- lane 5-7: TATB_INSB (PCR 3+4)
- lane 8-12: OmpRBS (PCR 6)
- lane 13: 1kb ladder
- lane 14-16: TATD_IDE (PCR 5+7)
- lane 17: 100bp ladder
- lane 18: OmpRBS (PCR 6)
re-culture pOmpC and pCinRHSL
- Because of the low concentration of pOmpC and pCinRHSL, the liquid culture is performed again to gain larger concentration
- concentrations
- pCinRHSL (0.05 ug/uL by pett, 0.05 and 0.08 by 海威)
- pOmpC #1 (run out off in previous experiment)
- pOmpC #2 (0.02 ug/uL by pett, 0.04 and 0.05 by 海威)
- pOmpC #3 (0.04 and 0.23 by 海威)
- pOmpC #4 (0.04 and 0.08 by 海威)
digestion for assembly
System 1
- pOmpC + TATA_INSA
- pOmpC (re-cultured, 10/2; S.P. digestion, 10/3)
- TATA_INSA (X.P. digestion, 10/2)
-
TATB_INSB + CinR+HSL+D-term- TATB_INSB (E.S. digestion, 10/2 (it is not correct!!! TATB_INSB does not have EcoRI site)
- CinR+HSL+D-term (E. digestion, 10/2)
- CinR+HSL+D-term (X. digestion, 10/3)
- (pOmpC + TATA_INSA) + (TATB_INSB + CinR+HSL+D-term)
- (pOmpC + TATA_INSA)
- (TATB_INSB + CinR+HSL+D-term)
- pOmpC + TATA_INSA + TATB_INSB
- pOmpC + TATA_INSA + TATB_INSB + CinR+HSL+D-term
System 2
- pCinRHSL + OmpRBS
- pCinRHSL (re-cultured, 10/2; S.P. digestion, 10/3)
- OmpRBS (X.P. digestion, 10/2)
- (pCinRHSL + OmpRBS) + TATD_IDE
- (pCinRHSL + OmpRBS)
- TATD_IDE
- [(pCinRHSL + OmpRBS) + TATD_IDE] + D-term
- [(pCinRHSL + OmpRBS) + TATD_IDE]
- D-term
