NYMU Taipei/Lab Notes/2007 10 4

From 2007.igem.org

(Difference between revisions)
(→Re-check the concentration of inserts and vectors)
(→ligation setup)
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</tr>
</tr>
</table>
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== ligation setup==
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== ligation setup ==
 +
* ligation I: pCinRHSL (vector) + OmpRBS (insert)
 +
<table border=1>
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<tr><td>pCinRHSL (vector)</td><td>3 uL</td></tr>
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<tr><td>OmpRBS (insert)</td><td>14 uL</td></tr>
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<tr><td>10X buffer</td><td>2 uL</td></tr>
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<tr><td>T4 ligase</td><td>1 uL</td></tr>
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</table>

Revision as of 14:48, 4 October 2007

digestion check of CinR+HSL+D-term,pCinRHSL and pOmpC (vectors)

NYMU Taipei 20071004 CinR-HSL-D-term,pCinRHSL,pOmpC digestion check.jpg
  • lane A: 1kb ladder
  • lane B: CinR+HSL+D-term x 2
  • lane C: 1kb ladder
  • lane D: pCinRHSL x 2
  • lane E: pOmpC x 2
  • after gel separation, their O.D. was checked. However, it is too low

Re-check the concentration of inserts and vectors

NYMU Taipei 20071004 3 vectors 3 inserts digestion check.jpg
  • lane A: 1kb ladder
  • lane B: pCinRHSL
  • lane C: pOmpC
  • lane D: CinR+HSL+D-term
  • lane E: 100bp ladder
  • lane F: TATA_INSA
  • lane G: TATB_INSB
  • lane H: OmpRBS
  • Owing to
    • concentrations of vectors are too low and
    • A260/A280 ratios are not correct (maybe too much ions)
  • thus, we checked the vectors with inserts by PCR to decide how to ligate them

ligation setup

  • ligation I: pCinRHSL (vector) + OmpRBS (insert)
pCinRHSL (vector)3 uL
OmpRBS (insert)14 uL
10X buffer2 uL
T4 ligase1 uL
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