Priya+A.

ToDo List:
Prepare RE Digest Samples for Middle Schoolers Dilute M13 Forward Ready Made Primer from IDT company Start looking at Overlap PCR cloning and start on Leishmania target

070212 - Priya, how did your pmCherry PCR turnout? If good - go onto pLIC PCR and then Overlap PCRs for your target.

062112 - Priya, try to figure out how to embed your images into the page instead of just having it link to the file. Also, add a REALLY good caption. - Thanks, DR. B

07.16.12: Annealing and Transformation

07.13.12: Made accepting vector and PCR inserts

07.11.12: Did PCR cleanup on second PCR^2

07.10.12: Did second PCR^2 and ran on gel

07.09.12: PCR Cleanup on PCR^2

07.06.12: Ran PCR Squared on gel Lane 1:PCR^2 Lane 2:PCR^2 Lane 3:PCR^2 Lane 4:PCR^2 Lane 7: 1kb DNA marker 07.05.12: PCR Squared

07.04.12: Secondary PCR; ran both primary and secondary on gel Lane 1: Skip Lane 2: 100 bp DNA Ladder Lane 3: 1kb DNA Ladder Lane 4: Primary PCR (PA) Lane 5: Secondary PCR Option B (PA) Lane 6: Skip Lane 7: Primary PCR, old Oligo Mix A (KR) Lane 8: Primary PCR, new Oligo Mix B (KR) Lane 9: Primary PCR, new Oligo Mix C (KR) Lane 10: Skip

07.03.12: Ran pLIC PCR on gel and started overlap PCR (primary PCR) Lane 2: 100bp DNA marker Lane 3: Tube A Lane 4: Tube B Lane 5: Tube C Lane 6: Tube D (no DNA control)

07.02.12: Ran pmCherry PCR on gel and did pLIC PCR Results

06.29.12: Worked on primer design and did pmCherry PCR

06.28.12: Worked on primer design

06.27.12: Worked on primer design

06.26.12: Prepared RE digest for middle schoolers, diluted M13For, Leishmania target

06.25.12: PCR Gel

Lane 4: 100bp DNA ladder Lane 5: Tube B- 1:1000 template dilution (3ng plasmid) Lane 6: Tube C- 1:100 template dilution (30 ng plasmid) Lane 7: Tube A- 1:1000 template dilution (0.3ng plasmid) Lane 8: Tube D- No DNA control

06.22.12: PCR (PGBR22)

06.21.12: Midi Prep with pmcherry and DH5alpha Nanodrop:

06.20.12: Beer's Law

06.19.12: PyMOL 1 and RE Digest Redo Lane 2: 1kb ladder Lane 4: Uncut DNA Lane 5: EcoRI Lane 6: PvuII Lane 7: EcoRI and PvuII This gel redo did not turn out as expected either. Some of the DNA got stuck in two of the wells, and there may be some contamination in lane 6.

06.18.12: Transformation Efficiency (pmcherry with DH5alpha) 1ng plasmid- approximately 960 colonies/ng

5ng plasmid- approximately 1008 colonies/ng

25ng plasmid- approximately 1108 colonies/ng

06.14.12, 06.15.12: RE Digest The bands did not appear as expected; the 4th and 5th lanes are probably contaminated, and the 6th lane does not have the spacing I expected. My virtual (expected) gel is below:

06.12.12: Micropipettor calibration

06.12.12: Submitting DNA for sequencing; Analysis: