Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/03/11: Difference between revisions
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Rene M Davis (talk | contribs) (Autocreate 2013/03/11 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs) |
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==Entry title== | ==Entry title== | ||
came in around 10am to check plates. some grew, picked colonies and grew up in lb amp | |||
ligations didn't have colonies yet | |||
came in a 7PM | |||
here's what grew from what was picked this morning: | |||
{| {{Table}} | |||
|- | |||
|1-2-1|| no | |||
|- | |||
|1-2-2|| no | |||
|- | |||
|2-2-1 ||no | |||
|- | |||
|2-2-2|| yes | |||
|- | |||
|4-2-1|| yes | |||
|- | |||
|4-2-2 ||yes | |||
|- | |||
|7-2-1 ||yes | |||
|- | |||
|7-2-2 ||no | |||
|- | |||
|10-2-1|| yes | |||
|- | |||
|10-2-2 ||no | |||
|- | |||
|12-1-1 ||yes | |||
|- | |||
|12-1-2 ||no | |||
|- | |||
|} | |||
some of the plates have more colonies. will run colony PCR to see what might be positive if these mini preps don't look good. | |||
<br> | |||
cut mini preps | |||
2 X & P | |||
yesterday's ligation: | |||
{| {{table}} | |||
|- | |||
|Part||colonies | |||
|- | |||
|Pconst bb control || 3 colonies | |||
|- | |||
|Pconst + RBS + C0061|| >20 colonies | |||
|- | |||
|Pconst + RBS + C0078|| 4-6 colonies (had low yield on gel extraction) | |||
|- | |||
|Pconst RFP bb control|| 3 colonies | |||
|- | |||
|Pconst RFP + K081023 || 2 colonies | |||
|- | |||
|} | |||
picked more colonies from ligations and parts i pulled out yesterday | |||
transformed some of the mini preps from today into BL21 | |||
2ul of mini prep into 10ul water | |||
added 20ul BL21 cells | |||
incubated on ice for 8-10 minutes | |||
heat shocked at 42deg for 45sec | |||
iced for 2 minutes | |||
plated | |||
K082035 (2-2-2) | |||
K084007 (4-2-1 and 4-2-2) | |||
K145254 (7-2-1) | |||
Revision as of 13:49, 28 May 2013
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Entry titlecame in around 10am to check plates. some grew, picked colonies and grew up in lb amp
ligations didn't have colonies yet came in a 7PM here's what grew from what was picked this morning:
some of the plates have more colonies. will run colony PCR to see what might be positive if these mini preps don't look good.
2 X & P
picked more colonies from ligations and parts i pulled out yesterday transformed some of the mini preps from today into BL21 2ul of mini prep into 10ul water added 20ul BL21 cells incubated on ice for 8-10 minutes heat shocked at 42deg for 45sec iced for 2 minutes plated K082035 (2-2-2) K084007 (4-2-1 and 4-2-2) K145254 (7-2-1)
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