Day 18 thoughts.  Phagefinders. 

Planned:

Chris: will make HT sample 19 F2 today. 

Did: 

Steve:  He compared the F2 S17 phage to F1 S17 phage, and noticed that F1 has an extremely pronounced halo that F2 does not have.  His proposal includes these 2 possibilities: 1) different smegmatis = different plaques.  2) room Temperature growth affects plaque size and type.  The bigger plaques are older, however they were grown for 24 h in the incubator and a weekend at room temperature. 

Therefore, he wil harvest S17 F2 as a HT phage today, and also continue studies with this phage by picking an F2 plaque (little plaque) and dilution plating this same plaque at either RT or at 37°C. 

S8b, is F1, and it will be picked for F2.

S8a, is possibly still contaminated with S8b, therefore, regardless, Steve will be pick and dilution plate another S8a (the one that is not B today.)

Steve needs to chart his history of plating, and if he is still seeing multiple phage plaque morphologies from individually picked plaques.  I'm curious if we are seeing multiple phages.  This should be a visible illustration of each sample, using arrows to describe when and where prepared sampmles contained more than 1 plaque or phage as visualized by EM.    

Genevieve: we will photograph the original garden plate that she has, and a plate that has been contaminated after picking from for better plaque morphology.  She plated 3 samples yesterday, today is just one day out and therefore it is too soon to tell.  The rain sample, no phage.  Not sure if she has 2 separate phages.  The plaques can be two separate sizes, we're not sure if big plaques are suppressed by smaller plaques. 

1A had been picked for F2 and plated yesterday; since Genevieve is waiting on 3 separate samples today, it is a photography and writing day for her. 

Today, Genevieve can dilution plate from her HT, and compare plaque morphologies at RT vrs. 37C.  10-4 from a plaque gives Genevieve about 268 plaques.  Expect that her HT phage is 10^10. 

Genevieve's question: why was the Mycobacteria paper (cell) put together in the manner that it was composed?  1) what manner it was put together in, 2) why

Lazlo:  we want to photograph the series of his phage, since when diluted, we see various plaque morphologies.  This is a nice demonstration of how different plaque shapes do not necessarily dilute as expected...it can be confusing to know if you have more than one phage species. 

Hypothesis: Lazlo is working with the same phage; he has a plate that he can make F2 HT from, tomorrow.  Therefore he is done picking, and needs to be hunting for another phage. He will start 2 new Central Park samples today, Beacon paragraph regarding their finds will also be discussed today.  Also, clone

Chris: had plated dilutions for F2 creek sample, which has evolved nice large plaques on the original plate.  These plaques are now quite small,

He can make a lacy plate from his 10-3,

we need to photograph his creek sample today, his BG sample has been HT made from F2, I think it is a good idea to dilution plate your HT and let these samples sit at either RT or at 37C, in order to determine if the phage has a temperature effect. 

Chris's question: describe location from where phages come from, the nature of their plaque morphologies, what you saw in the EM, if you think these phage will have related genomic sequence. 

Amanda:  has 2 different phages that come from either NC or NJ, and the morphologies are very similar.  NC phage is F1, so it needs be picked again today and dilution plated for a lacy plate and another plate that will verify the solution had pure phage in it.  Since last time, a plaque did not yield very many phages at -4 and -5, she only needs to plate -1, -2, -3. 

NJ phages is also F1, so iti needs to be picked today and dilution plated as above.  All plates today will then be left at RT for storage, and plates that have been picked from should be parafilmed and stored in the fridge. 

Chiara:  W100 sample from sidewalk, new phage is F0, it will be picked for the first time today, and this sample can also be photographed.  Her F1 from Canada can also be photographed.  Yesterday, Chiara did dilutions of CKCA (lucy), she picked from this and plated 2 separate sets of dilutions.  These will be F2.  The first set of dilutions 0-->-3, is at RT, the other is at 37 C. 

CKCA is really interesting, and is a very lysogenic and will be photographed so that it will go to the website.  (we need to talk later, more).  Chiara can also do the same Temperature based study with her other phage. 

Chiara is writing the Beacon paragraph that will state than new mycobacteriophages have been discovered by beacon students.  two sentence description regarding why this is significant.  Outstanding, scientifically interested sophomores and juniors should contast ..

Today's materials:   

Pointers:

If a student doesn't have a background in biology, and other students do, the students with the biology background are often really excited to teach the biology newcomer about the central dogma, DNA structure, and basic genetics.  It is a good learning exercise to encourage and give the students time for these activities.