Team:Panama/14 October 2010

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Prepared Ampicillin stock (50 mg/ml)
Prepared Ampicillin stock (50 mg/ml)
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We did a miniprep on the 11 colonies that grew in liquid LB medium with Amp. (The method we used was without columns because the kit ran out).
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Then, we ran a gel of 1% agarose to see if we could see a difference in the size of the linear band of the plasmid to be able to determine if it contains the insert (Rh1AB) or not.
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We did a miniprep on the 13 colonies that grew in liquid LB medium with Amp. (The method we used was without columns because the kit ran out).
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[[Image:Cuadro14octubre.JPG|500px|thumb|left|alt text]]
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Then, we ran a gel of 1% agarose to see if we could see a difference in the size of the linear band of the plasmid to be able to determine if it contains the insert (Rh1ab) or not.  
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'''Miniprep Protocol:'''
 
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1. Inoculate 3 ml LB medium (containing antibiotic) with a bacterial clone, culture with vigorous shaking at 37 degrees for 14-16 hours.
 
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2. Aliquot 2 x 750 ul culture into microcentrifuge tube, harvest bacteria by spinning at 12000rpm for 1 minute. Aspirate supernatent. Add an additional 750 ul culture media, respin and aspiate supernatant.
 
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3. Resuspend bacterial pellet by complete vortexing in 110ml resuspension buffer (100 ul solution A (25 nM Tris-HCl, pH 8.0, 10 mM EDTA) + 10 ul RNase A). The bacteria should be completely resuspended- no clumps should be visible. Solution A is stored in the refrigerator. RNAse is in the -30C freezer.
 
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4. Add 100 ul freshly prepared lysis buffer (50 ul 400 mM NaOH, 50 ul 2% SDS) and mix gently by
 
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*Photos
 
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[[Image:UntitledOct14_2.JPG|300px|thumb|left|alt text]]
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'''Miniprep Protocol:'''
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For this procedure we used what is called a "dirty miniprep". And by dirty we don't mean that we did it with unclean utensils nor did we do it after coming from a soccer game; it means that the reagents were prepared in the lab and they weren't obtained from a commercial kit.
{{:Team:Panama/calendar2}}
{{:Team:Panama/calendar2}}

Latest revision as of 03:49, 28 October 2010

iGEM Panama

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October 14

Prepared Ampicillin stock (50 mg/ml)

We did a miniprep on the 11 colonies that grew in liquid LB medium with Amp. (The method we used was without columns because the kit ran out). Then, we ran a gel of 1% agarose to see if we could see a difference in the size of the linear band of the plasmid to be able to determine if it contains the insert (Rh1AB) or not.

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Miniprep Protocol: For this procedure we used what is called a "dirty miniprep". And by dirty we don't mean that we did it with unclean utensils nor did we do it after coming from a soccer game; it means that the reagents were prepared in the lab and they weren't obtained from a commercial kit.

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Retrieved from "http://2010.igem.org/Team:Panama/14_October_2010"