--Justin Wang 14:57, 7 February 2011 (EST)
--Justin Wang 12:40, 15 February 2011 (EST)
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)
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)
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)
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)
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)
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)
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)
Sent for sequencing!
-edit our group confirmed all were correct
--Justin Wang 10:46, 10 March 2011 (EST)