2014hs.igem.org/team:Charlottesville RS/Notebook/041414

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<a href="https://2014hs.igem.org/Team:Charlottesville_RS/Parts" style="color: white">Parts<!--[if gt IE 6]><!--></a><!--<![endif]-->
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''Alli Ambrosini''
''Alli Ambrosini''
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The results of our transformation efficiency kit and John’s ligation thing were ready today, and they were very different. There was pretty much nothing on the efficiency kit, but the ligation results were very good. We got competent cells! We think there are a whole lot of reasons that our part didn’t work in the transformation efficiency kit, including possibly:
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The results of the transformation efficiency kit and John’s ligation procedure were ready today, and they were very different. There was no growth, but the ligation results were very good. The cells were competent. There were are several possibilities as to why our part did not work in the transformation efficiency kit, including:
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- We messed up in the beginning and never had any DNA at all
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- There was no DNA to begin with due to a mistake in procedure
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- Lauren and Alli messed up the volumes while pipetting during the lab
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- John’s group just might have gotten lucky and got all the competent cells, since we took them all from the same tube
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- The DNA wasn’t thawed enough, and we might not have picked up any
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'''Results:'''
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- Lauren and Alli incorrectly measured the volumes while pipetting during the lab- John’s group just might have gotten lucky and got all the competent cells, since they were all taken from the same tube
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-DNA/Chlor. -Transformation control = growth
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- The DNA was not thawed enough, and might not have picked up any DNA.  
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RFP Antibiotic Resistance
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-Parts A & B from iGem -Ligation control = growth
 
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-Transformation Efficiency Kit -Our part = no growth :(
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Results:
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'''For This Week:'''
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-DNA/Chlor. :
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transformation control = growth
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-Parts A & B from iGEM :
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ligation control = growth
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-Transformation Efficiency Kit :
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our part = no growth
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For This Week:
1. Rehydrate DNA and transform into E.Coli to multiply DNA
1. Rehydrate DNA and transform into E.Coli to multiply DNA
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2. Miniprep - rip open E.Coli to get purified DNA
 
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3. Digest - cut up the parts we need
 
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4. Ligate - stick em back together
 
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5. Transform (into giant robots)
 
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2. Miniprep
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3. Digest
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4. Ligate
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5. Transform
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Took bacteria from John’s “NEB10 beta” plate from 4/8/14 and streaked it onto the SOB agar plate in hopes of getting more bacteria.
Took bacteria from John’s “NEB10 beta” plate from 4/8/14 and streaked it onto the SOB agar plate in hopes of getting more bacteria.
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[[https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview Back]]

Latest revision as of 14:00, 20 June 2014

April 14th, 2014

Alli Ambrosini

The results of the transformation efficiency kit and John’s ligation procedure were ready today, and they were very different. There was no growth, but the ligation results were very good. The cells were competent. There were are several possibilities as to why our part did not work in the transformation efficiency kit, including:

- There was no DNA to begin with due to a mistake in procedure

- Lauren and Alli incorrectly measured the volumes while pipetting during the lab- John’s group just might have gotten lucky and got all the competent cells, since they were all taken from the same tube

- The DNA was not thawed enough, and might not have picked up any DNA.


Results:

-DNA/Chlor. : transformation control = growth

-Parts A & B from iGEM : ligation control = growth

-Transformation Efficiency Kit : our part = no growth


For This Week:

1. Rehydrate DNA and transform into E.Coli to multiply DNA

2. Miniprep

3. Digest

4. Ligate

5. Transform


Alli Ambrosini, Lauren Ewell

Took bacteria from John’s “NEB10 beta” plate from 4/8/14 and streaked it onto the SOB agar plate in hopes of getting more bacteria.

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