Transcription of Making Electrocompetent Cells
{{id}} {{{paragraph}}}
Example: bachelor of science
Making Electrocompetent Cells
Making Electrocompetent Cells Day 1. 1. Streak out frozen glycerol stock of bacterial Cells (Top 10, DH5 , etc.) onto an LB. plate (no antibiotics). Grow plate overnight at 37 C. Day 2. 1. Autoclave: 2 L of ddH2O. 100 mL of 10% v/v glycerol (molecular biology grade). 1 L LB (or your preferred media). 4 centrifuge bottles and caps Lots of microfuge tubes 2. Chill overnight at 4 C: ddH2O. 10% glycerol Centrifuge rotor 3. Prepare starter culture of Cells Select a single colony of E. coli from fresh LB plate and inoculate a 10 mL starter culture of LB (or your preferred media). Grow culture at 37 C in shaker overnight. Notes: You can also substitute other media like SOB, 2xYT, etc for LB if preferred.
6. Add 10ml of 1M MgCl2 7. Add 10ml of 1M MgSO4 8. Adjust pH to 7.0 with 10N NaOH and adjust volume to 1 L with distilled H2O. 9. Autoclave to sterilize on LIQUID cycle 15 minutes. 2xYT Medium 1. Measure ~900ml of distilled H2O. 2. Add 16g Bacto Tryptone.
Loading..
Tags:
Information
Domain:
Source:
Link to this page:
Please notify us if you found a problem with this document: