Transformation Protocol: Difference between revisions
Jump to navigation
Jump to search
>YanWu No edit summary |
>YanWu No edit summary |
||
Line 6: | Line 6: | ||
#Rescue cells with 500 uL SOC per tube | #Rescue cells with 500 uL SOC per tube | ||
#Shake at 37 C for 1 hr in incubator | #Shake at 37 C for 1 hr in incubator | ||
#Spread up to 20 uL on at least 2 plates for each library | #Spread up to 20 uL on at least 2 plates for each library. For control spread 100 uL on each plate, using at least 2 plates. | ||
#Add all tubes into large culture flask (150 mL) grow overnight | #Add all tubes into large culture flask (150 mL) grow overnight |
Revision as of 23:05, 6 May 2015
- Ensure water bath at 42 C, warm SOC media in incubator at 37 C
- Add 200 ng DNA into each tube of 100 uL cells (1-5 uL vector). You want to use up all of your vector so as to get the best coverage.
- Incubate on ice for 30 minutes
- Heat-shock for 45 seconds @ 42 C
- Incubate on ice again for 2 minutes
- Rescue cells with 500 uL SOC per tube
- Shake at 37 C for 1 hr in incubator
- Spread up to 20 uL on at least 2 plates for each library. For control spread 100 uL on each plate, using at least 2 plates.
- Add all tubes into large culture flask (150 mL) grow overnight