Talk:IGEM:MIT/2006/Communications: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 4: Line 4:


==Questions for Stephen==
==Questions for Stephen==
#Did you use the Knight or Endy site directed mutagenesis protocol? (http://openwetware.org/wiki/IGEM:MIT/2006/Notebook/2006-8-10#Mutagenesis says Knight but you told me Endy)
#*A: We used the Knight protocol.
#**Then are you sure that you phosphorylated the primers?  The Knight protocol doesn't call for this step.
*I'm sure we did.
#Do we have the regulated wintergreen GC data in a form that we could generate plots similar to the banana timecourse?
#Do we have the regulated wintergreen GC data in a form that we could generate plots similar to the banana timecourse?
*Yes, we could.  It would take some digging through old files and sorting out through each protocol we used on each day.  I don't know if it's necessary to show though.
#*Yes, we could.  It would take some digging through old files and sorting out through each protocol we used on each day.  I don't know if it's necessary to show though.
#What did you use to shake the cultures?  i.e. what incubator or shaker?
#What did you use to shake the cultures?  i.e. what incubator or shaker?
 
#*220 RPM- the shaker in 37 C room
220 RPM- the shaker in 37 C room
#*110 RPM- the shaker by Samantha's/2007 iGEM team's bench
 
110 RPM- the shaker by Samantha's/2007 iGEM team's bench


==General outstanding issues==
==General outstanding issues==

Revision as of 14:21, 11 February 2008


Questions for Stephen

  1. Do we have the regulated wintergreen GC data in a form that we could generate plots similar to the banana timecourse?
    • Yes, we could. It would take some digging through old files and sorting out through each protocol we used on each day. I don't know if it's necessary to show though.
  2. What did you use to shake the cultures? i.e. what incubator or shaker?
    • 220 RPM- the shaker in 37 C room
    • 110 RPM- the shaker by Samantha's/2007 iGEM team's bench

General outstanding issues

  • Funding sources for each person
  1. See last email... all undergrads were funded in the summer of 2006 by the MIT UROP office
  • Protocol for transformation

See below

  1. Transformation - André Transform competent cells
    • Thaw ~50 μl cells (whichever you like) on ice. Do not use glass tubes, which adsorb DNA.
    • Add 200 uL DNA to cells
    • Incubate on ice for 30 minutes
    • Incubate cells for 50-60 seconds at 42°C.
    • Incubate cells on ice for 2 min.
    • Add 200-300μL of room temperature SOC (not critical)
    • Incubate for 1 hour at 37°C on shaker.
    • Spread 200 μl onto plates made with appropriate antibiotic.
    • Grow overnight at 37°C.
  • Describing figure 6
  • Update <bbpart>BBa_Q04401</bbpart>
Retrieved from "https://openwetware.org/mediawiki/index.php?title=Talk:IGEM:MIT/2006/Communications&oldid=185709"