SBB11Ntbk-Justin Wang

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--Justin Wang 14:57, 7 February 2011 (EST)


First Post!


--Justin Wang 12:40, 15 February 2011 (EST)


Contents

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 picImage:022111-AnalGel3.jpg

   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)

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