Team:Caltech/Week 9

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
  • All colonies failed
• 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

• Ligation results:
  • Success: L31, L34, L77, L78, L79, L80
  • <partinfo>I15010</partinfo> and L33 both had < 5 colonies
  • Failed: L32, L76, L41, L60, L61
• Ran gel of yesterday's CPCR results: no significant bands
• More CPCR
  • 3x of each: L31, L34, L77, L78, L79, L80, L33, L58, L43

Sunday 8/15