User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/05/23: Difference between revisions
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#Cut to separate gel from bottom lip. | #Cut to separate gel from bottom lip. | ||
#Flip over and transfer gel to staining tray that has been prefilled with 100mL deionized water (see below). | #Flip over and transfer gel to staining tray that has been prefilled with 100mL deionized water (see below). | ||
# | #Add 100mL deionized water in staining tray. | ||
#Shake staining tray for 5 min on orbital shaker at room temperature. | |||
#Discard water. | |||
#Repeat steps 1-3 two more times. | |||
# | #Add 20mL SimplyBlue SafeStain (enough to just cover the gel). | ||
# | #Shake staining tray for 1 hr on orbital shaker at room temperature. | ||
# | #*''Overnight is better for improved sensitivity.'' | ||
#Repeat steps 9, 10 and 11. | |||
#Shake staining tray for 1 hr or more on orbital shaker at room temperature. | |||
#*''If the gel has been stained overnight, let it destain for a few hours.'' | |||
===Marker Sizes=== | ===Marker Sizes=== |
Revision as of 16:13, 28 May 2012
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5/23/2012Materials
ProcedureRunning buffer preparationDo this step first.
Sample preparation
Set up the gel apparatus during the 10 mins sample heating step. Note that prestained molecular weight marker doesn't need any preparation. After the heating process keep the samples on the bench to cool down to the room temperature. Running the gel
Staining the gel
Marker Sizes
Notes
Safety
See here for detailed safety information. References |