SBB11Ntbk-Justin Wang

--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.

New gel pic 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)