Difference between revisions of "User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/07"

From OpenWetWare
Jump to: navigation, search
(01/07/13)
(01/07/13)
Line 9: Line 9:
 
<!-- Precede finished items with a checkmark &#x2713; -->
 
<!-- Precede finished items with a checkmark &#x2713; -->
 
* Gibson re-transformation: grow cultures
 
* Gibson re-transformation: grow cultures
 +
* Re-transformation: 1x Gibson for pSB1A3 vector assemblies (3 & 4)
  
  
  
 
----
 
----
'''Gibson Re-transformation Results'''<br>
+
'''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
 
* 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.
 
* 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.
 
* Grow 2x 2 mL cultures from plate 3 streak to investigate whether an insert replaced RFP.
 +
 +
* Do Zymo minipreps
 +
 +
----
 +
'''Gibson Re-transformation'''<br>
 +
# hPCD + BL01 + pSB1A3 (#3), 1xGibson, '''6.0 μL'''
 +
# pSB1A3 (#4), 1xGibson, '''6.0 μL'''
 +
 +
* Add to 50 μL cells; ice/ 10 min.
 +
* Plate on 100 μg/mL amp LB agar
  
  

Revision as of 17:36, 7 January 2013

Owwnotebook icon.pngKarmella's BioBrick Cloning <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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