NYMU Taipei/Lab Notes/2007 10 2

From 2007.igem.org

< NYMU Taipei/Lab Notes(Difference between revisions)
(→System 1)
 
(23 intermediate revisions not shown)
Line 1: Line 1:
-
[[Image:NYMU Taipei TATA INSA INSB and OmpRBS GEL check.jpg|300px]]
+
== TATA_INSA, TATB_INSB, TATD_IDE and OmpRBS GEL check after PCR ==
 +
[[Image:NYMU Taipei TATA INSA INSB and OmpRBS GEL check.jpg|600px]]
[[Image:NYMU Taipei TATD IDE and OmpRBS GEL check.jpg|300px]]
[[Image:NYMU Taipei TATD IDE and OmpRBS GEL check.jpg|300px]]
 +
* 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''')
 +
* <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

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)
  • 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
Retrieved from "http://2007.igem.org/wiki/index.php/NYMU_Taipei/Lab_Notes/2007_10_2"