Haynes Lab:Notebook/Lab Traning - Pradyumna Kadambi

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(Pradyumna Kadambi/Undergraduate Lab Traning)
(Pradyumna Kadambi/Undergraduate Lab Traning)
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==Pradyumna Kadambi/Undergraduate Lab Traning==
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==Project Description==
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This notebook consists of the procedures learnt during lab training. DH5α will be transformed with KAH013 plasmid, and then miniprep will be performed, and a spectrophotometer will be used to analyze the DNA yield, and if the process was successful.
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'''DH5α Transformation with KAH013 Plasmid 6/25/2013'''<br>
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#In ice bath, 1μl KAH013 plasmid and 50μl competent E.Coli were combined.
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#Waited 10 minutes for bacteria to accept plasmid.
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#Transformed bacteria were then added to Agarose gel petri dish, rolling beads were used to evenly spread bacteria.
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#Petri dish was labeled with initials date and plasmid name("PK 6/25/2013 KAH13") and was placed in incubator at 37 degrees for two days(6/25-6/26).
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#Resultant colonies were picked by Rene and grown in liquid LB media overnight(6/27/2013).
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'''DH5α Incubation in Liquid Media 6/28/2013'''<br>
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#With a pipetter, 3mL of LB+amp was added to a test tube.
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#A colony was selected on the culture plate(plate was overgrown), and a circle was drawn where the colony was selected.
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#Micropipette tip was lightly touched to the colony making sure not to scrape any of the agar gel. Micropipette tip was then ejected into test tube.
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#Test tube was incubated with shaker for 7 hours at 37 degrees.
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''''DH5α Mini Prep with KAH013 6/28/2013'''<br>
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*Colonies that Rene had incubated in liquid media on 6/27 were used.
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#Using a micropipette, a 2mL centrifuge tube was filled with the incubated liquid media.
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#The solution was centrifuged at 13.1g for 3 minutes (If a pellet has not completely condensed centrifuge again until pellet does condense).
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#Using a micropipette, the supernatant was removed until 600μL of solution remained in the centrifuge tube. (Discard supernatant into the bleach container in the fume hood)
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#Pellet was broken up using the vortex.
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#Added 100μL of lysis buffer to the centrifuge tube and inverted 4-6 times and waited 1 min.
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#Added 350μL of lysis buffer. Invert the solution until it turns yellow, make sure no blue pockets are left.
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#Centrifuge the solution at
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Revision as of 22:10, 2 July 2013

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Project Description

This notebook consists of the procedures learnt during lab training. DH5α will be transformed with KAH013 plasmid, and then miniprep will be performed, and a spectrophotometer will be used to analyze the DNA yield, and if the process was successful.

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