r/Virology

Are there any Phage related conferences coming up this year? Or even other conferences in bacterial defense or phage anti defense?

i failed miserably to extract dna from my bacteriophages. i had 59 phages against pseudomonas aeruginosa isolates and 44 phages against salmonella enterica and 4 against enteroccous faecalis and 4 against e.coli bacteria. from high titre lysate i first tried to extract dna but the concentration was too less for sequecing. then i tried concentration using centrifugal filtration where i got reasoably good concentration for sequencing. 20-30% of my phages are sequenced and 70% are not sequenced. i am almost done with research but i am curious what went wrong from my side. also i feel i was a bad researcher.

i used standard plaque assay, selected plates with confluent lysis add 2-3ml of phage buffer, keep it in shaker for 30 minutes and scraped off top agar with phage buffer and transferred to centrifuge tubes and centrifuged at 3900g at 4 degree C for 15 minutes. supernatant was filtered using 0.22micron filter PES(Polyethersulfone) Membrane. this filtrate was concentrated using amicon centrifugal filter KDa molecular weight cutoff ultra centrifugal filter tubes. this concentrate was used for dna extraction.

kits used for DNA extraction - DNeasy blood and tissue kit, Qiagen. 100ul of high titer phage lysate + 75ul PBS. to this 20ul of DNase buffer and 5ul of DNase 1 were added and incubated at 37 degree Celcisu for 1 hour to remove host dna. then 180ul of ATL and 20ul of proteinase K were added and incubated at 57 degree C for 30 minutes. additional 20ul of proteinase K was added and incubated at 37 degree C and incubated over night. Day 2 200ul of AL buffer +200ul of absolute ethanol were added. vortexed for 30 seconds. this solution was transferred into kit column assembly folowed by washing steps and DNA was eluted in 50ul nuclease free water. eluted dna was estimated using nanodrop and qubit 4 platforms.

So, I learned that, the way the immune system handles viruses is that they would recognize them the first encounter, then after you survived and not die from said virus like they already made a wanted poster for the virus you won't get infected again by it?? Like chicken pox.

It's also how vaccines are made right? A weaker model of the virus is injected so that the immune system would go "I understand it now"

Then I recently learned that, the one responsible that makes us sick from time to time like the common cold is the rhinovirus, a virus.

So why can't the immune system recognize it?

Is there a research paper for this I can't find it.