Team:Caltech/Week 9

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Latest revision as of 16:16, 17 October 2010

iGEM 2010

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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

<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

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

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>
AIBN crosslinking ability test with linseed oil and hydrolized linseed oil.
- 3%-wt of AIBN mixed with 20ml of solution
- React for 5 hours at 60 degress Celsius water bath.
- Solution left at room at room temperature overnight
- white rod-shaped crystals were found in the solution second day
- the crystals are suspected to be products of AIBN self-reaction
- no sign of crosslinking

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

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@@ Line 72: / Line 72: @@
 * 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>
+*AIBN crosslinking ability test with linseed oil and hydrolized linseed oil.
+**3%-wt of AIBN mixed with 20ml of solution
+**React for 5 hours at 60 degress Celsius water bath.
+**Solution left at room at room temperature overnight
+**white rod-shaped crystals were found in the solution second day
+**the crystals are suspected to be products of AIBN self-reaction
+**no sign of crosslinking
 ==Weekend 8/14-8/15==
 === Saturday 8/14 ===
+* Ligation results:
+** Success: L31, L34, L77, L78, L79, L80
-=== Sunday 8/15 ===
+** <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
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