Minnesota/4 July 2008

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|'''[[Team:Minnesota/NotebookComparator|  Back to Notebook Home]]'''
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|'''[[Minnesota/3 July 2008|Go to Previous Day (July 3)]]'''|| width=158|'''[[Minnesota/7 July 2008|Go to Next Day (July 7)]]'''
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|1. '''PCR (Polymerase Chain Reaction):''' PCR used to modify Lambda_cI by copying a section of the gene, adding RBS, and using RS (restriction site enzyme) to cut copied gene section at the appropriate restriction site.  
|1. '''PCR (Polymerase Chain Reaction):''' PCR used to modify Lambda_cI by copying a section of the gene, adding RBS, and using RS (restriction site enzyme) to cut copied gene section at the appropriate restriction site.  
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::'''PCR Components in 50uL sln'''
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'''PCR Components in 50uL sln'''
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!Sequence Name !!Forward Primer !!Reverse Primer
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!| Components !!Volume
!| Components !!Volume
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|H20  ||35.0uL
|H20  ||35.0uL
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|2. '''Run PCR in Gel Electrophoresis:'''
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|a. Used 0.8% agarose gel, TAE buffer, and ethidium bromide (interculating agent)
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|b. Gel proved have correct Lambda_cI gene with 750 base pairs. Can now perform PCR purification (add buffers).
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|3. '''Miniprep MCherry gene'''
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|4.  '''Spectrophotometry:''' Used to determine the concentration of DNA in solution. Spec'd Lambda_cI with RBS (PCR sample).
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Latest revision as of 17:19, 7 July 2008

Back to Notebook Home
Go to Previous Day (July 3)Go to Next Day (July 7)
1. PCR (Polymerase Chain Reaction): PCR used to modify Lambda_cI by copying a section of the gene, adding RBS, and using RS (restriction site enzyme) to cut copied gene section at the appropriate restriction site. PCR Components in 50uL sln
Components Volume
Phusion HF Buffer 10.0uL
10 mM dNTP's 1.0uL
Primer mix 2.5uL
Template DNA 1.0uL
Phusion DNA Polymerase 0.5uL
H20 35.0uL



2. Run PCR in Gel Electrophoresis:
a. Used 0.8% agarose gel, TAE buffer, and ethidium bromide (interculating agent)
b. Gel proved have correct Lambda_cI gene with 750 base pairs. Can now perform PCR purification (add buffers).
3. Miniprep MCherry gene
4. Spectrophotometry: Used to determine the concentration of DNA in solution. Spec'd Lambda_cI with RBS (PCR sample).
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