User:Tk/Notebook/MF-xfm/2008/04/11

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 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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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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