Difference between revisions of "Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/03/11"

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(Autocreate 2013/03/11 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
 
(Entry title)
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==Entry title==
 
==Entry title==
* Insert content here...
+
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 12:49, 28 May 2013

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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:

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.
cut mini preps

2 X & P


yesterday's ligation:

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)