User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/07

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(01/07/13)
 
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* Gibson re-transformation: grow cultures
* Gibson re-transformation: grow cultures
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* Re-transformation: 1x Gibson for pSB1A3 vector assemblies (3 & 4)
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'''Gibson Re-transformation Results'''<br>
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'''Gibson Re-transformation Results (1/06/13)'''<br>
* Number of colonies did not improve. Only 1 colony on hPCD + BL01 + V0120 plate
* Number of colonies did not improve. Only 1 colony on hPCD + BL01 + V0120 plate
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* Streaks from yesterday:  
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* Streaks from yesterday: streak from plate 3 (hPCD + BL01 + pSB1A3 1xGibson) showed no color, whereas streak from plate 7 (hPCD + BL01 + pSB1A3 1/4x Gibson), showed RFP expression. The pSB1A3 template carries RFP.
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* Grow 2x 2 mL cultures from plate 3 streak to investigate whether an insert replaced RFP.
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* Do Zymo minipreps
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----
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'''Gibson Re-transformation'''<br>
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# hPCD + BL01 + pSB1A3 (#3), 1xGibson, '''6.0 μL'''
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# pSB1A3 (#4), 1xGibson, '''6.0 μL'''
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* Add to 50 μL cells; ice/ 10 min.
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* Plate on 100 μg/mL amp LB agar

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Karmella's BioBrick Cloning Main project page
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01/07/13

  • Gibson re-transformation: grow cultures
  • Re-transformation: 1x Gibson for pSB1A3 vector assemblies (3 & 4)



Gibson Re-transformation Results (1/06/13)

  • Number of colonies did not improve. Only 1 colony on hPCD + BL01 + V0120 plate
  • Streaks from yesterday: streak from plate 3 (hPCD + BL01 + pSB1A3 1xGibson) showed no color, whereas streak from plate 7 (hPCD + BL01 + pSB1A3 1/4x Gibson), showed RFP expression. The pSB1A3 template carries RFP.
  • Grow 2x 2 mL cultures from plate 3 streak to investigate whether an insert replaced RFP.
  • Do Zymo minipreps

Gibson Re-transformation

  1. hPCD + BL01 + pSB1A3 (#3), 1xGibson, 6.0 μL
  2. pSB1A3 (#4), 1xGibson, 6.0 μL
  • Add to 50 μL cells; ice/ 10 min.
  • Plate on 100 μg/mL amp LB agar



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