Day 24 thoughts. Phagefinders.
Planned: testing if boiling will kill phages and sterilize syringes. Also, will this sterilize filters?
Jeevan is coming in to talk to us about his thesis research. (Endolysin.)
Blake, how is your web-page?
Chiara, how is your web-page?
Possibilities are that low-titer DNA meant no phage DNA...need to make new high-titer DNA, soon.
Students will have time to set up their restriction digestions today, but will not have enough time to run their own samples on a gel. If there is time at the end of the day, students can add loading buffer to the cut DNA, and I will run the samples on the gel and produce photographs and digital images for them to look at.
Blake, please cast a gel.
AM: freezer boxes for DNA and phage, top agar for rhodo and myco, 7h9-agar into plates, petri-plates.
Did:
Restriction digestion will be done in 20 uL samples.
Regarding DNA preparation, Blake had good results using 1 mL 10^9 PFU High Titer phage preparations. His DNA had been suspended into 100 uL of water at the end of the procedure, and he would use 5 uL of this DNA for restriction digestion. We will take the same approach, except that I want to see more cut DNA. Each 20 uL cut will contain:
2 uL RE 10 x buffer, x 20 reactions. = 40 uL
0.2 uL acetylated BSA, x 20 = 4.0 uL
0.5 uL Enzyme, x 20 = 10 uL (Today, will do BSTeII)
10 uL phage DNA,
7 uL deionized H2O. x 20 =146 uL
The digestion will occur at 37°C for 2-3 hours, and will then immediately be loaded onto a gel. During the 2-3 hour gap. The student should have time to prepare new DNA today so that digestions can be done tomorrow with that DNA...the reason for having 2 digestion days.
To make master mix, we will mix for 20 cuts.
Chris rodococcus samples for rhodococcus phage hunting: 39, 27, 8, 3, 19, from the environmental sample library.
We discussed the cell paper further today. Students contemplated the value of bioninformatics as a result of the
Jeevan did not come in, therefore Chiara moderated a for or against, using debate procedures of the UN, whether the US should have a national bioinformatics system.
Moderated caucus, means when called on person has 30 sec to speak, called on by moderater. At the UN, it would be put on until someone puts on a motion to close the speakers list. I instead asked for back and forth. Everyone was put on the speakers list.
Lazlo: plan for med info access at any time within the comunity hospital, prevent leaking out, hackers from getting hold on it and black market for the information. Lazlo believes the hospital should have an internal server that holds the info in each hospital.
Kristen: I want you to picture this--you in a car, knocked out. only i.d. for you...allows for access of your info anywhere in the nation so that drs can help you. info flows with you, anywhere. probl is security issues... technology can be secure. CIA style. yet info can be accessed quickly.
Geneveeve: i understand value of history avail anytime, anywhere, but people's choice pro and cons ; pro=med histroy, cons= lose privacy. SS#'s get stolen, mine did. I have 2 now. if this can be stolen so easily, what about your med history? Pblms with security already. Main problem is that it is also hard to fund all this. Save our $.
Blake: history avail will save lives, but also will mean anyone with a good computer can get it. No way for pure security. Someone will take adv. of this info. Internet is a problem. Any skilled hacker. Blake then requested for an unmoderated cacus. Chiara said that a motion can be made to shorten/lengthen times. Chiara then kept a 2 min unmoderated caucus. They discussed further. Some
Chris: mentioned that a bioinformatics system was something that people could choose whether or not to do. Also, even if the information does leak out, the problems are not deadly. Insurance companies can be told not to look at information, this can be made an illegal deed. The information can not be life threatening in any way. Chris also plated the rhodococcus samples in order to attempt to find a phage capable of attacking this deadly disease.
Today's materials:
Pointers: