CRESMET Anna Hicks July 14-18

Monday - I prepared solutions to test for petri dish bacterial assay in the classroom.  Specifically, I needed to know if we could use the microwave instead of the autoclave to set the medium in the petri dishes.  The solution set perfectly using the microwave which will save me from having to drive down to ASU to autoclave the petri dishes in order to perform the lab.  I then took bacterial swabs of the doorknob and the elevator button to see if there was any bacterial/yeast growth.  The purpose was two-fold:  firstly to see if bacteria would reproduce on the microwaved agar and secondly, to see if it would duplicate appropriately without the use of the incubator as we do not have that option at the high school.  Tomorrow will tell us the answers.

Tuesday - There is almost no visible growth on the plates today.  Tomorrow I will check them again.  I checked chemical and equipment order books for catalog numbers and prices on all of the items needed for the plate assay and the ELISA.  Most were found but it took a long time to find the best prices for the ordering.  I retrieved the medium from the autoclave for the research project.

Wednesday - Today there is visible growth on the plates!  That's really a relief...but obviously room temperature will need more time to show the growth on the plates when the students perform this test at the school.  I cleaned equipment (the yucky job!) but got it mostly done.  We then (the entire lab staff) got to sit in on a Master thesis being defended!  Everything new and different is exciting!  This was interesting.  We then went back to the lab duties. 

Thursday - There is no lab this morning as our grad student has a medical appointment.

Friday - We will go in and see if there is anything to finish in the lab.

MSTF 2008 Hicks 4th week Protein HIV

Monday, July 7, Melissa is on vacation this week and we had decided previously that Trudi and I would use that time to compile our powerpoint, poster and lessons plans.  

Tuesday, July 8, we worked on our poster layout and categories.

Wednesday, July 9, we worked on verbage and state standards.

Thursday, July 10, we worked on picture layout and colors.

Friday, July 11, I had an appointment this morning.

MSTF 2008 Hicks 3rd week Protein HIV

Monday, June 30, Melissa and Dr. Ghirlanda examined the phage colonies growing in the petri dishes and neither were satisfied with what they were seeing.  In a review of protocol and procedures there was no obvious indication as to just why and at what point in the procedure something changed from the original.  It was decided that Melissa would call the company after returning from vacation on Monday, July 14 and see if there was any suggestion that they would make about the material which they supplied.

Tuesday, July 1, we spent reading material and cleaning up some of the equipment in the lab.

Wednesday, July 2, we went over some of the equipment we would need to perform a lab at our high school.  We checked the order book to check prices on some of the equipment to see what would be feasible to order for the high school.

Thursday, July 3, Trudi and I started working on the lesson for the high school and putting together the pictures for the powerpoint presentation.   

MSTF 2008 Hicks 2nd week Protein HIV

Monday we spent most of the time in lab reading different material.

Tuesday we spent mixing 3 different solutions for the research assignment (this will probably be a weekly necessity for each week's assay). We mixed top agarose, M9TB and LB. All mixtures were then brought to the proper pH and autoclaved. (This took the entire four hours although it should have been accomplished quicker, however, this was the first time that Trudi and I were actually performing the task ourselves.)

Wednesday was spent becoming familiar with a testing procedure called ELISA. Dr. Ghirlanda has supplied us with the particular protocol for this test and explained the procedures involved in the ELISA. This particular test is well documented for lab tests performing titers assaying antigens against specific antibodies. We will be starting the ELISA tomorrow to experience the procedures and results obtained by this particular method of testing.

Thursday we became familiar with ELISA procedures and the washing of the protein, pipetting, and diluting the serum. Dr. Ghirlanda guided us through the different steps actually showing how the 96 cells are filled with protein and subsequently washed and subjected to antibody solution. We used a multiple pipette applicator for the first time and could see how specific laboratory instruments are crucial to performing experiments in a timely fashion.

Friday we worked with Melissa and picked up where she stopped on Thursday afternoon.  We viewed gel electrophoresis and she showed us how she determined the ligation of the DNA strand.  We also looked at the photo density of the solution to determine the correct photo density before proceeding.  Melissa had already worked on preparing the phage plates so they were ready for a final count.  The plates did not look like the original so she chose to put them aside until Dr. Ghirlanda could see them.

Protein binding of specific cell membrane site resulting in inhibition of HIV virus entry using E. coli

Our work is being led by Giovanni Ghirlanda, the principal investigator and Melissa Ruben, grad student.

Monday we were shown the location of most of the lab work for this project, given the first set of papers regarding the project to read and met Melissa.

Tuesday we were shown the correct concentration of material and liquids required to produce the medium required for the growth of the e. coli virus. In doing this, correct sterility, non-contamination lab skills and lab safety (i.e. gloves, close-toed shoes, googles and common lab sense) were covered. We subsequently mixed both medium and agar solution and adjusted pH to 7.5 before we autoclaved both mixtures. Proper use of the autoclave, measuring instruments, sterility of lab equipment (using bleach for E. coli containers), use of the pipette calibration unit, and the optical density calibrator were shown.

Wednesday we were given the second paper on their lab project to read and study. Melissa had prepared petri dishes with E. coli cultures the afternoon before and we examined the number of phage circles present on each.

Thursday we met Giovanna Ghirlanda and Melissa took us to the chemistry computer lab to show us the chemistry program available to the chemistry department here at ASU. Great stuff!

Friday Giovanna gave us the short version of the protein binding project and clarified a few grey areas for us. Melissa showed us how to use the DNA concentrator and prepared a new mixture for electrophoresis.