Research+Page-+Christina

**Christina's Research**
Christina - show some images (screenshot of BLAST result) - DR. B


 * Week 10: **
 * (show image or data, not just summary or what you did) - Dr. B **
 * Finished Characterization
 * Concentrated Protein
 * FPLC
 * Concentrated FPLC product


 * Week 9: **
 * Finished Expression
 * Finished Purification
 * Began Characterization

**Week 8:**
 * Mini Prep Colonies
 * Send to DNA sequencing
 * Received Results and Blasted them
 * Colony 6 worked


 * Began Expression with Transformation into BL21(DE3)


 * Week 7: **
 * Received new primer and diluted it
 * Restarted Cloning
 * PCR off of pNIC-Bsa4 with the old gene sequence in it


 * Cut accepting vector (pNIC-Bsa4)


 * Cohesive End Generation
 * Annealing and Transformation
 * Overnight Growth

**Week 6:** Christina - ok good deal. Don't bother including .docx links here - just keep those in your Google Docs. But rather show the most critical information of your results here (either data or images like the one below)
 * Began Protein Purification
 * Made SDS-Page Gels
 * Began Protein Characterization
 * Did not see our protein in the Elutions
 * Decided that it was because our protein had a trans-membrane section encoded by the first few residues
 * Confirmed this using TMPRED
 * Ordered new Custom Forward Primers


 * Week 5: **


 * Received results from sequencing.
 * After blasting the results, both forward and reverse sequences matched the FrTuHP gene.
 * Successful cloning of FrTuHP gene into pNIC-Bsa 4
 * Began Protein Expression


 * Week 4: **


 * Received results from sequencing.
 * After blasting the results all the forward sequences matched the FrTuHP gene except the last 200 bp were missing.
 * After also blasting the reverse sequences both colonies 2 and 3 matched all of the FrTuHP gene without any missing base pairs.
 * Grew up colony 2 overnight in 80mL of LB
 * Spun down and Midi prep the colonies.
 * Sent those results to sequencing with forward and reverse primers.
 * Completed Virtual Screening Refresher


 * Week 3: **


 * PCR amplification of FrTuHP from pUC19 (scaled up version)
 * Cut pNIC-Bsa 4 accepting vector
 * Cohesive end generation of both accepting vector and PCR insert
 * Annealing and Transformation
 * Overnight growth of 4 colonies and Miniprep of those colonies
 * Sent Miniprep results to sequencing with colonies 1, 2, 3, and 4 having forward primers and 2 and 3 also having reverse primers.
 * Completed PyMOL refresher


 * Week 2: **


 * Received results from sequencing but the results contained long sections of N's in the middle of the sequence.
 * Restarted pNIC-Bsa 4 cloning.

**Week 1:**


 * Mini prep pNIC-Bsa 4 cloning colonies that were spun down over the summer.
 * Sent Mini prep results to DNA sequencing

**Transformation Efficiency**

[[image:SAM_0382.JPG width="800" height="600" caption="Transformation of 25ng of pGBR22 plasmid with transformation efficiency of 192,000"]]





 * RE Digest Gel **

[[image:RPKPpGBR22gel.JPG width="800" height="607" caption="Restriction Enzyme Digest Gel of pGFP. Lane 2: Ladder. Lane 5: EcoRI. Lane 7: PvuII. Lane 8: EcoRI + PvuII. Lane 9: uncut pGFP "]]

 * PCR #1 pGBR22 **



**PCR #2 GREEN**