Transformations
- still no transformants
- E. coli control worked well
- 1.25 KV control gave 100 transformants (1.75 hr. outgrowth)
- 1.75 KV control gave 850 transformants (1.75 hr. outgrowth)
- tried again today with 50 ml culture spun down to 250 ul with 2x wash in EPBG
- culture was sligntly turbid, yellow
- Four transformations:
- plain
- with 1 ul BSA added (NEB 100x)
- with 1 ul pUC19 DNA added (1 ug/ul)
- with both BSA + DNA added
- electroporation at 1.75 KV, 1 mm gap
- outgrowth 1.5 hours
- no sparking
- cultures turned yellow
- plated 300 ul
- also redid E. coli control
Possible problems
- bad plates -- test growth of older MF resistant strains on the new plates
- plated earliest cultures A,B,C and wt MF on a tet plate (4/9/08)
- temperature bad?
- but normal MF cells grow fine
- cell concentration still too low
- obviously much lower than E. coli dense cell growth
- should also test trypsin treatment
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MF-xfm
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