Protocol of transformation:
Prepare the HeatBlock and shaker before getting XL1 cells
1. Put water into the blocks
2. Turn on the heatblock and set the temperature at 65C
3. Put a thermometer in the heatblock to track the temperature
4. Turn on the shaker, 250 rpm, 37C
Start the Trasnformation
1. Take out XL1 cells from -80 freezer and thaw on ice
*The ones we use in the lab have 50ul of cells/tube
*The ones we use in the lab have 50ul of cells/tube
*Make sure to take out some spares in case there's not enough cells
2. Take out SOC and the same time and thaw at room temperature
*If you need to use an old one, make sure it looks clear
3. Prepare some 2ml tubes: one tube for each transformation and label the tube with the name of the plasmid
4. Set the pippetors ready:
One P200 at 115ul for XL1 cells
One P2 at 2ul for plasmids
One P1000 at 500ul for SOC
5. Open some XL1 cells and leave the tubes on ice
6. Mix tubes of XL1 cells by pipetting
*Do not centrifuge XL1 cells since they are fragile
*If there’re big drops of cells on the edge, use the pippet
tips to get it down to the bottom
*If there’re small drops of cells on the edge, just leave it
7. From the mixed tube, take out 115ul of XL1 cells and add to
the marked 2ml tubes
Leave the tubes on ice after added XL1 cells
8. Add 2ul of plasmids to
115ul of XL1 cells
9. Leave the 2ml tubes in
fridge for 30 minutes
10. Take the tubes out
11. Heat shock at 42C for
45 seconds in HeatBlock
12. Take the tubes out
13. Leave the tubes in
fridge for 2 minutes
14. Take the tubes out
15. In the hood, add 500ul
SOC into the cells
*Drop the SOC onto the cells nice and slow
*No need to mix afterwards
16. Leave the tubes in the
shaker for 4 hours
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