Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/04/29

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(Autocreate 2014/04/29 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
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Ran PCR products from yesterday on a gel. 5ul of reactions 2-9, all 30 of 1 so I can gel extract because there should be two bands and I only want one.  
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<br>
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DpnI digest PCR reactions 2-7 and 9 with positive and negative controls (didn't run reaction 8 because I checked the gel early and saw that there was no DNA there).
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<br>
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Controls
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Positive control was a miniprepped plasmid at 100ng/ul concentration.
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|'''Volume (ul)'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Thing||Positive ctrl||Negative ctrl
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|-
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| H2O||12||12
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|-
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| Green digest buffer||2||2
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|-
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| pTet mCh||5||5
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|-
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| DpnI||1||0
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|}
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<br><br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Thing'''
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| align="center" style="background:#f0f0f0;"|'''Volume (ul)'''
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|-
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| Digest buffer||4
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|-
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| PCR rxn||25
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|-
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|H2O||1
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|-
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| DpnI||1
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|}
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Incubated at 37°C for 55 minutes, sat on the bench for 3 minutes, heat inactivated at 80 for 5 minutes.<br>
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Cleaned using zymo DNA clean and concentration-5 kit (columns from gel extraction kit). Added 150ul of DNA binding buffer to DpnI reactions.
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<br>
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Gel extracted PCR reaction 1. Saw a smaller band above, think I got the correct band.
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Revision as of 15:03, 29 April 2014

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mm/dd/yyyy

Ran PCR products from yesterday on a gel. 5ul of reactions 2-9, all 30 of 1 so I can gel extract because there should be two bands and I only want one.
DpnI digest PCR reactions 2-7 and 9 with positive and negative controls (didn't run reaction 8 because I checked the gel early and saw that there was no DNA there).
Controls Positive control was a miniprepped plasmid at 100ng/ul concentration.

' Volume (ul) '
ThingPositive ctrlNegative ctrl
H2O1212
Green digest buffer22
pTet mCh55
DpnI10



Thing Volume (ul)
Digest buffer4
PCR rxn25
H2O1
DpnI1

Incubated at 37°C for 55 minutes, sat on the bench for 3 minutes, heat inactivated at 80 for 5 minutes.
Cleaned using zymo DNA clean and concentration-5 kit (columns from gel extraction kit). Added 150ul of DNA binding buffer to DpnI reactions.
Gel extracted PCR reaction 1. Saw a smaller band above, think I got the correct band.


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