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==
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* Insert content here...
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came in around 10am to check plates. some grew, picked colonies and grew up in lb amp
 +
 
 +
 
 +
 
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ligations didn't have colonies yet
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came in a 7PM
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 +
here's what grew from what was picked this morning:
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{| {{Table}}
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|-
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|1-2-1|| no
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|-
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|1-2-2|| no
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|-
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|2-2-1 ||no
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|-
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|2-2-2|| yes
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|-
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|4-2-1|| yes
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|-
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|4-2-2 ||yes
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|-
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|7-2-1 ||yes
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|-
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|7-2-2 ||no
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|-
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|10-2-1|| yes
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|-
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|10-2-2 ||no
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|-
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|12-1-1 ||yes
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|-
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|12-1-2 ||no
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|-
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|}
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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.
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<br>
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cut mini preps
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2 X & P
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 +
 
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yesterday's ligation:
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{| {{table}}
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|-
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|Part||colonies
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|-
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|Pconst bb control || 3 colonies
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|-
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|Pconst + RBS + C0061|| >20 colonies
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|-
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|Pconst + RBS + C0078|| 4-6 colonies (had low yield on gel extraction)
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|-
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|Pconst RFP bb control|| 3 colonies
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|-
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|Pconst RFP + K081023 || 2 colonies
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|-
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|}
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picked more colonies from ligations and parts i pulled out yesterday
 +
 
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transformed some of the mini preps from today into BL21
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2ul of mini prep into 10ul water
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added 20ul BL21 cells
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incubated on ice for 8-10 minutes
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heat shocked at 42deg for 45sec
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 +
iced for 2 minutes
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plated
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K082035 (2-2-2)
 +
 
 +
K084007 (4-2-1 and 4-2-2)
 +
 
 +
K145254 (7-2-1)
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Revision as of 15:49, 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:

Partcolonies
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)



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