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  The Ward lab at UC Santa Cruz

Strain maintenance

  • Get GE24 pha-1(e2123) III from CGC, make glycerol stocks

  • Maintain at 15ºC on NGM-lite plates seeded with HB101 (available from CGC).  If making a lot of knock-ins, I chunk to a fresh 10 cm plate every day, so that I have correctly staged animals at any given time.  I inject either young adults or day 1 adults with a nice row of eggs

  • One additional advantage of cku-80 RNAi (besides increased knock-in efficiency), is that it is easier to pick worms off of HT115 bacteria compared to HB101
cku-80 RNAi
Construct and oligo design
PU6::sgRNA templates by PCR fusion
Injections
FAQs
PDF of methods and consideration
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