Minnesota/4 July 2008

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Latest revision as of 17:19, 7 July 2008

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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. 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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+{|style="align:left" width="965"
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+|'''[[Team:Minnesota/NotebookComparator|  Back to Notebook Home]]'''
+|-
+|'''[[Minnesota/3 July 2008|Go to Previous Day (July 3)]]'''|| width=158|'''[[Minnesota/7 July 2008|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.
-|}
 {|
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 |-
 |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).
 |}