The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>
|
2/5/13
Bacterial transformation
- Added total volume (10.0 μL) to 50 μL chemically competent cells (e.g., BL21) in a 2.0 mL tube.
- Incubated on ice for 2 min., heat shock at 42°C for exactly 45 sec., and placed on ice.
- Added 800 μL sterile SOC medium.
- Grow with shaking at 37°C for 30 min.(Taped tubes to the shaker rack.)
- Pelleted the cells at top speed in a microcentrifuge for 3 min. at room temperature.
- Discarded the supernatant. Resuspended the cells in 100 μL LB + antibiotic.
- Plate cells on pre-warmed LB agar + antibiotic. (I plated 100 μL of the cells. Then put beads on plates and shook.) Protocol didn't specify.) Grow overnight at 37°C.
- Check results tomorrow.
|