Latest revision as of 18:47, 7 August 2008

Untitled Document

Today in the Lab

Tammy and Dan

Sample	Concentration 1 (ng/μL)	V1 (μL)	Concentration 2 (ng/μL)	V2 (μL)	H2O (μL)
T123	75	4	5	60	56

PCR Amplification of T123

Control for PCR Reaction was ddH2O instead of DNA template

12 PCR Reactions with identical DNA template

Matt

PCR cleanup

Performed a PCR cleanup of LiG ptp2/Pssa42

Transformation

Transformed Ligated PTP2/pSSA42 into competent cells.

Inoculation

Inoculated more pSSA42 vector for future digestion/ligations.

@@ Line 80: / Line 80: @@
-:<li> '''Digestion Step 1 of pSSA14 with PstI'''
+:<li> '''PCR Amplification of T123'''
 <TABLE BORDER=2>
-<TR> <TD>'''Digestion Reaction components'''</TD><TD>'''Vol (&mu;l)'''</TD> </TR>
+<TR> <TD>'''PCR Reaction components'''</TD><TD>'''1X Vol (&mu;l)'''</TD><TD>'''13X Vol (&mu;L)'''</TD> </TR>
-<TR> <TD>H2O</TD><TD>4</TD> </TR>
+<TR> <TD>5X Reaction Buffer</TD><TD>10</TD> <TD>130</TD></TR>
-<TR> <TD>Buffer 3</TD><TD>1.5</TD> </TR>
+<TR> <TD>10 mM each dNTP</TD><TD>1</TD><TD>13</TD> </TR>
-<TR> <TD>BSA</TD><TD>1.5</TD> </TR>
+<TR> <TD>F Primer (10 pmol/&mu;L) F69</TD><TD>2.5</TD><TD>32.5</TD> </TR>
-<TR> <TD>PstI</TD><TD>1</TD> </TR>
+<TR> <TD>R Primer (10 pmol/&mu;L) F70</TD><TD>2.5</TD><TD>32.5</TD> </TR>
-<TR> <TD>DNA template</TD><TD>7</TD> </TR>
+<TR> <TD>DNA Template</TD><TD>4</TD><TD>52</TD> </TR>
-<TR> <TD>'''Total'''</TD><TD>'''15'''</TD> </TR>
+<TR> <TD>Phusion Polymerase</TD><TD>0.5</TD><TD>6.5</TD> </TR>
+<TR> <TD>Filtered Sterile ddH2O</TD><TD>29.5</TD><TD>383.5</TD> </TR>
 </TABLE>
+::<li> Control for PCR Reaction was ddH2O instead of DNA template
+::<li> 12 PCR Reactions with identical DNA template
+'''Matt'''
+:'''PCR cleanup'''
+::<li> Performed a PCR cleanup of LiG ptp2/Pssa42
+:'''Transformation'''
+::<li> Transformed Ligated PTP2/pSSA42 into competent cells.
+:'''Inoculation'''
+::<li> Inoculated more pSSA42 vector for future digestion/ligations.