NYMU Taipei/Lab Notes/2007 10 2

From 2007.igem.org

(Difference between revisions)
(System 1)
(System 1)
Line 25: Line 25:
** pOmpC ('''re-cultured, 10/2'''; '''S.P. digestion, 10/3''')
** pOmpC ('''re-cultured, 10/2'''; '''S.P. digestion, 10/3''')
** TATA_INSA ('''X.P. digestion, 10/2''')
** TATA_INSA ('''X.P. digestion, 10/2''')
-
* TATB_INSB + CinR+HSL+D-term
+
* <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>''')
** 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 ('''E. digestion, 10/2''')
Line 31: Line 31:
* (pOmpC + TATA_INSA) + (TATB_INSB + CinR+HSL+D-term)
* (pOmpC + TATA_INSA) + (TATB_INSB + CinR+HSL+D-term)
** (pOmpC + TATA_INSA)
** (pOmpC + TATA_INSA)
-
** (TATB_INSB + CinR+HSL+D-term)
+
** (TATB_INSB + CinR+HSL+D-term)</s>
=== System 2 ===
=== System 2 ===

Revision as of 04:42, 9 October 2007

Contents

TATA_INSA, TATB_INSB, TATD_IDE and OmpRBS GEL check after PCR

NYMU Taipei TATA INSA INSB and OmpRBS GEL check.jpg NYMU Taipei TATD IDE and OmpRBS GEL check.jpg

  • 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)

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