User:Meng Xiao He/Notebook/fall08/2008/12/09: Difference between revisions
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[[Image:12-5_pcr_strips.jpg]] | [[Image:12-5_pcr_strips.jpg]] | ||
* unfortunately, EtBr seems to have run so far up gel that bands < 1kb can't be distinguished | * unfortunately, EtBr seems to have run so far up gel that bands < 1kb can't be distinguished | ||
** flecks are from bubbles | |||
** some of the first few samples didn't seem to have left well? | |||
* 1-5 were colonies 1, 3, 8, 9, and 10 from the 11-30 "new cco" KO | * 1-5 were colonies 1, 3, 8, 9, and 10 from the 11-30 "new cco" KO | ||
* then ladder | * then ladder | ||
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[[Image:12-5_pcr.png]] | [[Image:12-5_pcr.png]] | ||
=== | ===Remainder of samples with bands rerun at higher res=== | ||
[[Image:Hires.png]] | [[Image:Hires.png]] | ||
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====Conclusions==== | ====Conclusions==== | ||
* program used ('''12/8''' on big room machine) seems to work (at least some of the time) with the outer primer sets (in this case, all 3 were added) | * program used ('''12/8''' on big room machine) seems to work (at least some of the time) with the outer primer sets (in this case, all 3 were added) | ||
* cbb colonies of interest: new 2, new 3 | * cbb colonies of interest: [(new 2, new 3)→ambiguous b/c cannot see if the WT 5.4kb cbb outer fragment is indeed missing] | ||
* cco colonies of interest: new 1, new 2 | **new4, new 5 don't look like they incorporated the vector, since the WT 5.4kb cbb outer fragment is still there- what are those bands btw 1.6kb and 2kb? | ||
* cco colonies of interest: new 1, new 2 (a little iffy> is there still the 6.5 kb WT cco outer fragment?) | |||
==Insertional colony processing== | ==Insertional colony processing== | ||
* restreaked on LB Gm plates | * restreaked on LB Gm plates | ||
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* cco new # 1 (rename to cco12.9A) | * cco new # 1 (rename to cco12.9A) | ||
* cbb new # 3 (rename to cbb12.9A) | * cbb new # 3 (rename to cbb12.9A) | ||
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Revision as of 10:16, 12 December 2008
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12/7 Platinum PCR results1KB ladder
12/8 Phusion PCR resultsWithin each block (end with ladder): cco 3R > cco 5F > Duo 3R > Duo 5F > cbb 3R > cbb 5F Blocks go: (new filter colony 1 > 2 > 3 > 4 > 5 > 6) > old colony > wt control
Remainder of samples with bands rerun at higher res
Conclusions
Insertional colony processing
LB minus salt:
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