SBB11Ntbk-Justin Wang: Difference between revisions
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--[[User:Justin Wang|Justin Wang]] 14:57, 7 February 2011 (EST) | |||
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First Post! | First Post! | ||
--[[User:Justin Wang|Justin Wang]] 12:40, 15 February 2011 (EST) | |||
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<h2> Entry 1 </h2> | |||
Prepared my oligonucleotides for PCR. | |||
-followed protocol for sbb1110 and sbb1129 | |||
-Oligos are aleady diluted to 10 microliters | |||
-setup pcr solutions to be places. | |||
--[[User:Justin Wang|Justin Wang]] 12:40, 17 February 2011 (EST) | |||
<h2> Entry 2 </h2> | |||
<i>PCR Product Failed. There was no volume in the pcr product.</i> | |||
Redid the PCR setup following protocol. | |||
Set up the Eco/Bam digestion for the cut and paste process. jtk2259 | |||
-Used thermocycler to start the digestion | |||
-ran analytical gel on the part. Cut out the large bane in geland purified with zynos cleanup | |||
--[[User:Justin Wang|Justin Wang]] 12:40, 18 February 2011 (EST) | |||
<h2> Entry 3 </h2> | |||
Ran the Gel on the PCR products - sbb1110 and sbb1129 | |||
-Did the gel electrophoresis protocol. | |||
-analyzed gel picture | |||
-sbb1110 failed. No bands need redo. | |||
Zymos cleanup on the gel product of the jtk2259. | |||
--[[User:Justin Wang|Justin Wang]] 12:40, 21 February 2011 (EST) | |||
<h2> Entry 4 </h2> | |||
Experiment previously used wrong AW buffer and the entire previous process was redone. | |||
New gel pic[[Image:022111-AnalGel3.jpg]] | |||
sbb1110 lane 10 | |||
sbb1129 lane 12 | |||
Digested the product | |||
sbb1110 gel D pos 2 | |||
sbb1129 gel D pos 3 | |||
--[[User:Justin Wang|Justin Wang]] 10:46, 24 Feburary 2011 (EST) | |||
<h2> Entry 5 </h2> | |||
Zymos Cleanup on the digested PCR Products. | |||
Ran the GEL and cut out bands for the cut and paste product. | |||
--[[User:Justin Wang|Justin Wang]] 10:46, 1 March 2011 (EST) | |||
<h2> Entry 6 </h2> | |||
Performed ligation procedure on the PCR products. | |||
Heat shock transormation in to cells | |||
Plated cells on Kan antibiotics | |||
--[[User:Justin Wang|Justin Wang]] 10:46, 3 March 2011 (EST) | |||
<h2> Entry 7 </h2> | |||
Colonies were picked for us. | |||
Began miniprepping the cultures. | |||
4 colonies of each was cultured. | |||
1 sample of sbb1110 failed to turn purple. Everything else was good. | |||
--[[User:Justin Wang|Justin Wang]] 10:46, 8 March 2011 (EST) | |||
<h2> Entry 8 </h2> | |||
Sent for sequencing! | |||
-edit our group confirmed all were correct | |||
--[[User:Justin Wang|Justin Wang]] 10:46, 10 March 2011 (EST) |
Latest revision as of 00:53, 20 April 2011
--Justin Wang 14:57, 7 February 2011 (EST)
First Post!
--Justin Wang 12:40, 15 February 2011 (EST)
Entry 1
Prepared my oligonucleotides for PCR.
-followed protocol for sbb1110 and sbb1129 -Oligos are aleady diluted to 10 microliters -setup pcr solutions to be places.
--Justin Wang 12:40, 17 February 2011 (EST)
Entry 2
PCR Product Failed. There was no volume in the pcr product.
Redid the PCR setup following protocol.
Set up the Eco/Bam digestion for the cut and paste process. jtk2259
-Used thermocycler to start the digestion -ran analytical gel on the part. Cut out the large bane in geland purified with zynos cleanup
--Justin Wang 12:40, 18 February 2011 (EST)
Entry 3
Ran the Gel on the PCR products - sbb1110 and sbb1129
-Did the gel electrophoresis protocol. -analyzed gel picture -sbb1110 failed. No bands need redo.
Zymos cleanup on the gel product of the jtk2259.
--Justin Wang 12:40, 21 February 2011 (EST)
Entry 4
Experiment previously used wrong AW buffer and the entire previous process was redone.
sbb1110 lane 10 sbb1129 lane 12
Digested the product
sbb1110 gel D pos 2 sbb1129 gel D pos 3
--Justin Wang 10:46, 24 Feburary 2011 (EST)
Entry 5
Zymos Cleanup on the digested PCR Products.
Ran the GEL and cut out bands for the cut and paste product.
--Justin Wang 10:46, 1 March 2011 (EST)
Entry 6
Performed ligation procedure on the PCR products.
Heat shock transormation in to cells
Plated cells on Kan antibiotics
--Justin Wang 10:46, 3 March 2011 (EST)
Entry 7
Colonies were picked for us.
Began miniprepping the cultures.
4 colonies of each was cultured. 1 sample of sbb1110 failed to turn purple. Everything else was good.
--Justin Wang 10:46, 8 March 2011 (EST)
Entry 8
Sent for sequencing!
-edit our group confirmed all were correct
--Justin Wang 10:46, 10 March 2011 (EST)