Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/04/30

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(Autocreate 2014/04/30 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
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Purified PCR reactions 5 and 6 from yesterday (receiver backbone) using the gel extraction columns and the DNA clean and concentrator kit. Got really low yield, need to look into a better kit for this.  
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<br><br>
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Will set up golden gate with the purified PCR backbones. Will gel extract the other ones and then run those tomorrow if this doesn't work. Will only do one of each because I don't have enough receiver backbone.<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Description'''
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| align="center" style="background:#f0f0f0;"|'''ng/µL'''
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| align="center" style="background:#f0f0f0;"|'''length'''
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| align="center" style="background:#f0f0f0;"|'''fmol/ul'''
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| align="center" style="background:#f0f0f0;"|'''ul for 200 fmol'''
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|-
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| Rec bb||31||2203||21||9.52
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|-
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| pLux-GFP||47.148||1051||68||2.94
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|-
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| pRhl-GFP||50.466||1051||72||2.78
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|-
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| pEsa-GFP||32.1||1051||46||4.35
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|-
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| pLac-LuxR||45.771||817||89||2.25
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|-
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| pLac-RhlR||59.88||765||119||1.68
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|-
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| pLac-EsaR||37.847||758||74||2.70
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|}
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Reaction'''
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| align="center" style="background:#f0f0f0;"|'''Rec bb'''
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| align="center" style="background:#f0f0f0;"|'''pLux-GFP'''
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| align="center" style="background:#f0f0f0;"|'''pLac-LuxR'''
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| align="center" style="background:#f0f0f0;"|'''BsmBI'''
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| align="center" style="background:#f0f0f0;"|'''T4 ligase'''
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| align="center" style="background:#f0f0f0;"|'''T4 ligase buffer'''
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| align="center" style="background:#f0f0f0;"|'''H2O'''
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|-
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| Lux Rec||9.52||2.94||2.25||0.5||1.25||2||1.54
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|-
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| Rhl Rec||9.52||2.78||1.68||0.5||1.25||2||2.27
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|-
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| Esa Rec||9.52||4.35||2.7||0.5||1.25||2||-0.32
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|-
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| Bb control||9.52||0||0||0.5||1.25||2||6.73
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|}
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Revision as of 12:57, 30 April 2014

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mm/dd/yyyy

Purified PCR reactions 5 and 6 from yesterday (receiver backbone) using the gel extraction columns and the DNA clean and concentrator kit. Got really low yield, need to look into a better kit for this.

Will set up golden gate with the purified PCR backbones. Will gel extract the other ones and then run those tomorrow if this doesn't work. Will only do one of each because I don't have enough receiver backbone.

Description ng/µL length fmol/ul ul for 200 fmol
Rec bb312203219.52
pLux-GFP47.1481051682.94
pRhl-GFP50.4661051722.78
pEsa-GFP32.11051464.35
pLac-LuxR45.771817892.25
pLac-RhlR59.887651191.68
pLac-EsaR37.847758742.70


Reaction Rec bb pLux-GFP pLac-LuxR BsmBI T4 ligase T4 ligase buffer H2O
Lux Rec9.522.942.250.51.2521.54
Rhl Rec9.522.781.680.51.2522.27
Esa Rec9.524.352.70.51.252-0.32
Bb control9.52000.51.2526.73


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