Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2015/05/18: Difference between revisions

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==05//2015==
==05/18/2015==
<font color = purple>
I ran the PCR-optimization samples from last Friday on a gel; 4 were treated with DMSo and 4 were treated without (used water in place of it). The ones than were ran without DMSO appeared to have darker bands and lighter off-bands, so we decided to PCR purify those four. <br>
Without DMSO samples, pcr purified:
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Sample Read#'''
| align="center" style="background:#f0f0f0;"|'''260'''
| align="center" style="background:#f0f0f0;"|'''280'''
| align="center" style="background:#f0f0f0;"|'''260/280'''
| align="center" style="background:#f0f0f0;"|'''ng/µL'''
|-
| 1||0.014||0.005||2.72||14.471
|-
| 2||0.022||0.011||1.89||21.666
|-
| 3||0.011||0.005||2.082||10.794
|-
| 4||0.011||0.005||2.18||11.399
|}


We also fast-transformed AubI and BjaI from last Monday's ligations with Ryan's new cells (NEB 10β), because we have been seeing colonies on the negative control using René's cells. In addition to the two ligations, we made a negative control plate and a backbone-only control plate with the double digested EX sender backbone. <br><br>
Ryan ran a gel for two colonies on each of the plates we transformed on Monday, and of those, two colonies seem to have been successfully ligated (Lux1 and Rpa2 (?)).




Latest revision as of 00:57, 27 September 2017

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05/18/2015

I ran the PCR-optimization samples from last Friday on a gel; 4 were treated with DMSo and 4 were treated without (used water in place of it). The ones than were ran without DMSO appeared to have darker bands and lighter off-bands, so we decided to PCR purify those four.
Without DMSO samples, pcr purified:

Sample Read# 260 280 260/280 ng/µL
1 0.014 0.005 2.72 14.471
2 0.022 0.011 1.89 21.666
3 0.011 0.005 2.082 10.794
4 0.011 0.005 2.18 11.399

We also fast-transformed AubI and BjaI from last Monday's ligations with Ryan's new cells (NEB 10β), because we have been seeing colonies on the negative control using René's cells. In addition to the two ligations, we made a negative control plate and a backbone-only control plate with the double digested EX sender backbone.

Ryan ran a gel for two colonies on each of the plates we transformed on Monday, and of those, two colonies seem to have been successfully ligated (Lux1 and Rpa2 (?)).



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