Research+Page-+JC

Week 13:
=12/2/11= Inhibition Test with compound 7575772. (Marshall's Top Scoring Compound) =11/30/11=

Inhibition Test with compound 5153298
=11/29/11= =Vary Substrate (pNPP) Test= = = =11/28/11= Phosphatase Enzyme Test Enzyme Works

Week 12:
11/21 FLPC and Concentration. The size of the YopH protein is 33512.8 daltons.

Week 11:
=11/18/2011= Dried the gel in the biotech lab for 2 hours at 70 C. =11/15/2011= Ran all my samples on an SDS page gel. Gel shows that my protein sample might have some impurities.

Week 10:
=11/09/11= Got rid of pellet and stored supernatant in frdige. =11/08/11= Attended Symposium at AT&T Conference center. =11/07/11= Submitted top 5 compounds to order. Top compound was 5153298

Week 9:
=11/05/2011= Day 3 of Protein Expression. Stored pellet sample in -80 C freezer. Weight of Pellet = 2.78 g =11/04/2011= Day 2 of Protein Expression. =11/03/11= Ran some virtual screen jobs against the ChemBridge-Diversit3d.sdf library. Had to make several conf files since the library was too big to screen. =11/01/11= Positive results from DNA sequencing. Image coming later.Began Day 1 of expression. Transformed plasmid into BL21 cells. Grew up positive colony from master plate too.

Week 8:
=10/27/2011= Nanodropped and sent 4 samples with plic primers to sequencing.

=10/26/2011= Centrifuged colonies and miniprepped.

=10/24/2011= Prepared master plate. picked 8 colonies to grow up. =10/23/11= Cohesive end generation of PCR Inserts and Accepting Vector. Anealing and transformation.

Week 7:
= = =10/20/11= Submitted first run of the HF9 Library. No colonies grew up on plate. Prepared more vector. Made more PCR product. =10/19/11= Cohesive End Generation on PCR insert and Accepting Vector and Anealing and Transformation and gel check on pNIC-Bsa vector simultaneously. =10/16/11 Prepared pNIC-Bsa4 vector with BsaI.=

Week 6:
=10/14/2011=

YopH gene (921 bp) looks the correct size. =10/13/2011 Sequencing Results=

=10/10/2011= Sent 4 samples with for/rev primers to DNA sequencing

Week 5: 10/3/11
Completed Transformation on 10/07/11. DNA sequencing on Oct 10.

=Transformation on 10/4/11=

Week 4: 9/26/11
Was not able to log in a lot of hours this week. Will make up for it.

Week 3: 9/19/11
Prepared 3 more samples of Pnic Bsa4 vector. need to purify and gel check. Prepared accepting PNic Bsa4 vector. Failed



Week 2: 9/12/11
Little work done in lab. Spent a lot of time working on research poster and at LSAMP conference.

Week 1:
J.C. - looks good. - Dr. B

3rd Try: Primary/Secondary PCR Results 9/7/11
2ndary PCR doesn't look too good. Ran PCR squared after. Might have to run again for a better band or use some of marshall's sample.

2nd Try: Primary/Secondary PCR Results 9/6/11
Used old primary pcr from the first try and made a second primary pcr. Diluted Custom primers. Used first primary pcr sample for the third try because it was similar to the primary pcr sample from the summer.

pNIC- Bsa4 (CA7) Target: HSCD00041026
1st Try. Gel was to thin, so the image would not show up on screen. Didn't work though.