Talk:DIYbio:Demo: Difference between revisions
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# dip the tube into 42°C water for 1 minute (use the other container, the microwave, and a thermometer to heat the water. Be precise!) | # dip the tube into 42°C water for 1 minute (use the other container, the microwave, and a thermometer to heat the water. Be precise!) | ||
## put the tube back on ice for 2 minutes | ## put the tube back on ice for 2 minutes | ||
# add 500 | # add 500 μL of LBamp | ||
## tape the tube to a 37°C heat source, like the back of a computer for 1 hour. | ## tape the tube to a 37°C heat source, like the back of a computer for 1 hour. | ||
# Dump onto an agar+amp plate - or - dump into 5 mL of LBamp in a culture tube. | # Dump onto an agar+amp plate - or - dump into 5 mL of LBamp in a culture tube. | ||
Latest revision as of 23:19, 6 July 2008
Instructions for the basic 3-step transformation:
Preflight checklist:
- Box of competent cells on ice
- Part Datasheets (includes part dna)
- hole-punch tools (one per part)
- sterile Luria Broth + ampicillin (LBamp)
- 10 mL culture tubes
- disposable pipets (1mL)
- tape (for securing tubes to heat source)
- 2 containers (one of water bath, one for ice bath)
- microwave
- thermometer
- punch out the dna spot from the datasheet of the part you want to use
- and drop the "dna chad" into a thawed tube of competent cells
- and leave this tube on ice for 30 minutes
- dip the tube into 42°C water for 1 minute (use the other container, the microwave, and a thermometer to heat the water. Be precise!)
- put the tube back on ice for 2 minutes
- add 500 μL of LBamp
- tape the tube to a 37°C heat source, like the back of a computer for 1 hour.
- Dump onto an agar+amp plate - or - dump into 5 mL of LBamp in a culture tube.
- Incubate your plate or tube at the 37°C heat source overnight
see DIY placenta stem cells for graphic inspiration of this how-to
