SBB11Ntbk-RishiRawat: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Rishi Rawat (talk | contribs) No edit summary |
Rishi Rawat (talk | contribs) |
||
| Line 5: | Line 5: | ||
==[[User:Rishi Rawat|Rishi Rawat]] 12:58, 17 February 2011 (EST)== | ==[[User:Rishi Rawat|Rishi Rawat]] 12:58, 17 February 2011 (EST)== | ||
I messed up the PCR. I used 2.4uL H20 instead of 24uL. Will redo the PCR. | I messed up the PCR. I used 2.4uL H20 instead of 24uL. Will redo the PCR. | ||
The CORRECT mixture: | |||
24uL ddH2O | |||
3.3uL 10x Expand Buffer "2" | |||
3.3uL dNTPs (2mM in each) | |||
1uL Oligo 1, 10uM | |||
1uL Oligo 2, 10uM | |||
0.5uL Expand polymerase "1" | |||
0.5uL Template DNA | |||
==[[User:Rishi Rawat|Rishi Rawat]] 02:03, 15 February 2011 (EST)== | ==[[User:Rishi Rawat|Rishi Rawat]] 02:03, 15 February 2011 (EST)== | ||
Revision as of 11:00, 17 February 2011
~~!~~
Rishi Rawat 12:58, 17 February 2011 (EST)
I messed up the PCR. I used 2.4uL H20 instead of 24uL. Will redo the PCR.
The CORRECT mixture:
24uL ddH2O 3.3uL 10x Expand Buffer "2" 3.3uL dNTPs (2mM in each) 1uL Oligo 1, 10uM 1uL Oligo 2, 10uM 0.5uL Expand polymerase "1" 0.5uL Template DNA
Rishi Rawat 02:03, 15 February 2011 (EST)
Recieved oligo RRsb1118Forward, resuspended in water(added 295 uL ddH2O) to make 100uM stock.
Did PCR for sb1118, 1122, 1102 used mg1655 template
sb1118 F: sb1118F R: ss43R goal:371bp
sb1122: F: ss48F R: ss48R goal:1010bp
sb1102: F: ss27F R: ss27R goal:986bp
used extend protocol:
2.4uL h20 3.3uL 10x expand2 3.3uL dNTP 1uL oligo 1uL oligo .5uL expand .5uL Template
Placed reaction tubes in <2k bp PCR Reaction
Rishi Rawat 02:03, 9 February 2011 (EST)
Hello!
Rishi Rawat 02:02, 9 February 2011 (EST)
This is my first entry!
