Team:BYU Provo/Notebook/Phage Purification/Fallexp/Period1/Exp/10.25T4PCR

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| colspan="3" | <font color="#333399" size="5" font face="Calibri"> '''Phage Purification September - October Notebook: Experiments'''</font>

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: <u> '''Phage Purification''' </u> </font>

: [[Team:BYU Provo/Notebook/Phage_Purification/Winterexp|March-April]]

: [[Team:BYU Provo/Notebook/Phage_Purification/Springexp|May-June]]

: [[Team:BYU Provo/Notebook/Phage_Purification/Summerexp|July-August]]

: [[Team:BYU Provo/Notebook/Phage_Purification/Fallexp|September-October]]

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<font size="5"> '''10.25 T4 PCR''' </font>

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'''I) Purpose'''
: Identify DNA sequence difference in capsid proteins between mutants. 

'''II) Expected Outcome'''
: PCR products of the T4 wild type and mutant capsid protein genes.

'''III) Reagants Used'''
: 2 microL T4 wild type or mutant phages in each.
::TAQ PCR
: 2.5 microL Thermo Pol. buffer (for high fidelity TAQ)
: 1 microL primers  (B1 297/B1 298) (B1 302/B1 303)
: 17 microL ddH20
: 1 microL 10 mM dNTP's
: .5 microL Polymerase (TAQ)

TAQ was run to amplify and sequence the capsid proteins using two sets of different primers. (4 samples)

'''IV) Actual Procedure'''
: Put 2 microL phage in an eppendorf tube. Incubate at 99 C for 5 min. 
: Add the above reagents in the order listed to each tube.
: Run PCR using the TAQ protocol
: Run in 1% gel
:: 100 mL 1x TAE buffer
:: 1 g regular agarose
:: heat until dissolved completely
:: cool and add 2 drops ethidium bromide
:: pour with 14 well mold and allow to solidify
:: add 2 microliters of dye to 4 new tubes
:: extract 5 microliters of each phage and put them into each of the tubes.
:: We put the DNA ladder in well 1, well 2 was WT 297/298, well 3 was MUT 297/298, well 4 was WT 301/302, well 5 MUT 301/302

'''V) Results'''
:  
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[[File: | thumb|none|alt=A|T4 PCR Gel2]]



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