Day 13 thoughts.  Phagefinders. 

Planned:

discussion on where our contamination is.  discussion of our Crypto and Candida samples, and phage-hunting that was done therein.

student's given time to summarize their data and form their paragraphs and strategies.  We will monitor our smegmatis cultures that we started the day before. 

Mention samples left out, not in incubator, or not shaking overnight.  Reinforce that students need to be working on their paragraphs that describe what they need to be up to. 

questions for students: comprehensive paragraph describing results so far, using all appropriate identifiers (acronyms) and describing what samples we have collected.  Also, they should describe their strategy for the next few weeks regarding isolation of new phage. 

Amanda: Incubated 2 new soil samples yesterday, anything on the plates yet?  Wants to incubate additional samples? 

Kristen: will be able to harvest from essentially all of her samples, and will also be prepping more

bring in: 

Chris:  New and old smeg data table.  How is this? 

Lazlo:  Has 2 new samples to plate today. 

Stephen:  Do we have new plaques?  Pick and plate if so. 

Blake:  How did it go with isolating phage DNA? An advanced procedure. 

Reid:  prepare new top-agar for leaving; tomorrow it needs to be autoclaved early.  Check on the Smeg sample that I started yesterday. 

Genevieve:  Sit down with Genevieve, ensure all samples are properly labeled.  Genevieve's soil samples that incubated overnight need to be plated today. 

Did: 

Bill comes in at 10.  Since so much can be lost by not going over existing data, students were also given time to write up the data that they had to date. 

The following question was posed: In a concise paragraph or page, write about the samples that you have collected, and if you have phage or not.  Where did you get phage from?  What is unique about your plaque morpohology?  If you haven't collected phage yet, describe the samples that you have attempted to collect phage from.  Why do you suppose that you have not found a new phage yet?

Amanda: Incubated 2 new soil samples yesterday, too early to tell if anything on the plates.  Wants to incubate additional samples? 

Kristen: will be able to harvest from essentially all of her samples, and will also be prepping more ///  Kristen has some differences in what the quality of smeg did with respect to the phage plaque morphology.  See picture I took.  I'm actually not sure if it is the smeg that was different.  No contamination.  For 3BF1, 10-2 has visibly less plaques than 10-4.  There is a possiblity that this, even though picked from a plaque, the sample appears to contain 2 different phages.  I believe this because the larger colonies dilute from 10-1 unto 10-3.  Therefore, Kristen will pick a large plaque from 10-2, where the small plaques are not visible.  I think teh big guys repress the small guys.  The big guy is now going to be phage B.1.  Pick a second colony from the 10-5 plate, note that these all have turbidity except for the middle.  This other plaque is B.2.  The 10-4 was harvested.  It's label is 3BF1. 

3AF2 (original plate-->harvest-->plate again-->harvest-->plate again) is the colony that comes off the plate now.  It has been plated as much as it needs to be plated further.  A and C have been plated this much, already.  Even though A has been plated this much, we are concerned that a phage with a very small colony morphology is hitchiking with the other phages, as for sample B.  Therefore, The HT F1 from A will be dilution plated and we will see on Friday if we have solo phage.  (If found on Friday, into the fridge.  Regardless, the plates get left at RT then.) In addition, the A plate 10-2 has been set aside for possible lacy plate harvesting, if in fact we only have 1 phage in the dilution plating. 

C is very lytic, clear at 10-1, almost at 10-2, 10-3...

For the compost, we are harvesting the 10-1 and are picking a plaque from 10-3.   (Chiara continues here.)

Dilution plating the A from HT: plate -5 to -9. (Kristen continues here)

Dilution plating CH: plate 0 unto -4.  It had made few plaques at -3 this time.

Dilution plating of B:  Plate -2, -4, -6. (Kristen working with B.2) (Amanda works with B.1)

C to do: harvest the plate, this is purified enough and can be harvested as a lacy plate tomorrow.  Remaining -1, -2 that are not being harvested are left in the incubator to see if they will generate resistant phage.  Today, record colony # on C 10-1. 

Photograph B today. xx 

Again, it is possible that both A and B have same small phage. 

bring in: 

Chris:  New and old smeg data table.  How is this? 

Lazlo:  Has 2 new samples to plate today. 

Stephen:  Do we have new plaques?  Pick and plate for F2, Decided to plate to -5, from -1,-2,-3,-4,-5.  This is Sample #8. 

Blake:  How did it go with isolating phage DNA? An advanced procedure.  He is using Cla1.  Prabakaran screens with different enzymes.  Cla1

Chiara:  Has been partnered with Kristen today, too much going on there. 

Reid:  prepare new top-agar for leaving; tomorrow it needs to be autoclaved early.  Check on the Smeg sample that I started yesterday.  Get HT phage from people for EM. 

Genevieve:  Sit down with Genevieve, ensure all samples are properly labeled.  Genevieve's soil samples that incubated overnight need to be plated today. 

Today's materials:   

Pointers: