Difference between revisions of "Haynes Lab:Notebook/Engineering PC-TFs/2012/11/08"

From OpenWetWare
Jump to: navigation, search
(Autocreate 2012/11/08 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
 
(Summary)
Line 7: Line 7:
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
==Summary==
 
==Summary==
*  
+
Assembly Check of PCR Reactions 1,2,3,4<i>
 +
 
 +
* Gel Electrophoresis
 +
 
 +
Follow steps for gel electrophoresis.
 +
*Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray.
 +
*Create 1% gel by putting .6 grams of agarose into flask.
 +
*Microwave agarose solution for 30 seconds
 +
*Using hot gloves, grab flask out of microwave and swirl flask vigorously. Place back in microwave for 30 more seconds.
 +
*When flask is taken out of microwave, make sure that the agarose is completed dissolved.
 +
*Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the thin "teeth" and slip in the notches.
 +
*Add EtBr (ethidium bromide) to the agarose gel. There should be 1 microliter of EtBr per 10ml of agarose gel. Therefore, 6 µL.
 +
*Pour gel into tray. Wait twenty minutes for gel to settle.
 +
*Wash the agarose gel flask.
 +
*Pipette 15 μL of DNA ladder into first well. Pipette 5μL of PCR reactions into subsequent wells.
 +
*Turn on electrophoresis and let run for thirty minutes.
 +
 
 +
The assembly did not work. No bands were seen when placed under UV light.<b>
 +
 
  
  

Revision as of 17:15, 11 November 2012

Owwnotebook icon.png Engineering PC-TFs <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>

Summary

Assembly Check of PCR Reactions 1,2,3,4

  • Gel Electrophoresis

Follow steps for gel electrophoresis.

  • Orient top notch of gel electrophoresis apparatus. Place gel tray into larger tray.
  • Create 1% gel by putting .6 grams of agarose into flask.
  • Microwave agarose solution for 30 seconds
  • Using hot gloves, grab flask out of microwave and swirl flask vigorously. Place back in microwave for 30 more seconds.
  • When flask is taken out of microwave, make sure that the agarose is completed dissolved.
  • Set up wells on gel electrophoresis apparatus. The mold for the first digest is B-10. Use the thin "teeth" and slip in the notches.
  • Add EtBr (ethidium bromide) to the agarose gel. There should be 1 microliter of EtBr per 10ml of agarose gel. Therefore, 6 µL.
  • Pour gel into tray. Wait twenty minutes for gel to settle.
  • Wash the agarose gel flask.
  • Pipette 15 μL of DNA ladder into first well. Pipette 5μL of PCR reactions into subsequent wells.
  • Turn on electrophoresis and let run for thirty minutes.

The assembly did not work. No bands were seen when placed under UV light.