User:Tk/Notebook/MF-xfm/2008/04/11: Difference between revisions
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(Autocreate 2008/04/11 Entry for User:Tk/Notebook/MF-xfm) |
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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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Revision as of 21:47, 19 June 2008
 MF-xfm
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Transformations
Possible problems
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