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Monday 8/9
- Group meeting!
- Re-ran two gels of CPCR results
- Successful: 55-5, 56-1/-2/-3, 57-1/-2/-3, 54-5/-6
- Prepped DNA for use in digestions tomorrow
Tuesday 8/10
- Digested DNA for ligations:
- L31: L17 + <partinfo>K215000</partinfo> + Kan
- L32: L19 + L20 + Tet
- L33: L19 + L18 + Tet
- L34: L17 + L21 + Amp
- L35: L18 + L21 + Amp
- L41: L26 + <partinfo>B0015</partinfo> + Tet
- L49: HSP + L40 + Tet
- L60: <partinfo>K215000</partinfo> + L57 + Tet
- L61: <partinfo>K215000</partinfo> + L56 + Tet
- L63: L29 + L20 + Amp
- L64: L27 + L20 + Amp
- Prepped 37-1, 54-5/-6, 55-5, 56, 57 DNA
- Started CPCR of L36, L38
- Started liquid culture of <partinfo>I15008</partinfo>
Wednesday 8/11
- <partinfo>I15008</partinfo> culture did not grow; will re-pick more tomorrow.
- L43 plate had no colonies; will redo L43 tomorrow.
- Began CPCR of all successful ligations (31, 32, 33, 34, 35, 36, 38, 41, 49, 60, 61, 62, 63, 64).
- Gels:
- gel 1: 35-2 & 63-3 successful
- gel 2: 64-1 successful
- gel 3: 36-6 successful
- gel 4: 62-1 & 62-3 successful
Thursday 8/12
- Prepped 4 successful (non-red) cultures: 35-2, 36-6, 62-1, 63-3
- Digested:
- 35 (us)
- 36 (us)
- 37 (ds)
- 63 (us)
- Started cultures of 62-1, <partinfo>I15008</partinfo>, <partinfo>I15009</partinfo>
- Ligations:
- L76: L35 + L20 + Tet
- L58: L36 + L37 + Cm
- L43: L17 + HSP-amp + Tet
- CPCR: 31, 32, 33, 34, 38, 41, 49, 60, 61, 64
- gels appeared to have no bands at all... will re-run tomorrow.
- Transformed L76, L58, L43
Friday 8/13
- Ligation results:
- L76: 0 colonies; failed
- L58: > 500 colonies
- L43: ~100 colonies
- CPCR gels rerun from yesterday appear just as bad. 64-5 looks correct, but all other lanes are empty.
- Cultured L55-5
- Digested: <partinfo>I15008</partinfo> (ds), <partinfo>I15009</partinfo> (ds), L42 (bbone)
- Ligated:
- L41: L26 + <partinfo>B0015</partinfo> + T
- L60: <partinfo>K215000</partinfo> + L57 + T
- L61: <partinfo>K215000</partinfo> + L56 + T
- L76: L35 + L30 + T
- L77: L42 + <partinfo>pSB1C3</partinfo>
- L78: L44 + L64 + T
- L79: L19 + <partinfo>I15008</partinfo> + C
- L80: <partinfo>B0034</partinfo> + <partinfo>I15009</partinfo> + C
- CPCR: L76 x 6, L58 x 3; will run the gels tomorrow.
- Transformed: L31, L32, L33, L34, L41, L60, L61, L76, L77, L78, L79, L80, <partinfo>I15010</partinfo>
Weekend 8/14-8/15
Saturday 8/14
Sunday 8/15
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