M13 phage pfu determination - (Feb/07/2007 )
Hi,
I have obtained new M13 phage but I don't know how to calculate the number of plaque forming units/ml. I think I will need to infect bacterial cells with a known volume of the new M13 phage solution but I don't know how many cells should I infect and or plate in order to count the number of plaques or if i should mix this with medium-agar and pour it into a Petri dish.
I would be very grateful if someone could give me the protocol.
Thanks a lot in advance.
Rebeca
-RMG-
QUOTE (RMG @ Feb 7 2007, 10:57 AM)
Hi,
I have obtained new M13 phage but I don't know how to calculate the number of plaque forming units/ml. I think I will need to infect bacterial cells with a known volume of the new M13 phage solution but I don't know how many cells should I infect and or plate in order to count the number of plaques or if i should mix this with medium-agar and pour it into a Petri dish.
I would be very grateful if someone could give me the protocol.
Thanks a lot in advance.
Rebeca
I have obtained new M13 phage but I don't know how to calculate the number of plaque forming units/ml. I think I will need to infect bacterial cells with a known volume of the new M13 phage solution but I don't know how many cells should I infect and or plate in order to count the number of plaques or if i should mix this with medium-agar and pour it into a Petri dish.
I would be very grateful if someone could give me the protocol.
Thanks a lot in advance.
Rebeca
your M13 phage vector have LaCZ report gene or other report gene. I have calculated the tilter of phage in phage display. the general protocol: 1. mix the phage with host cell, and IPTG 2. spread mixture onto the plate 3. pull the low agar onto the plate 4. calculated blue plaque.
-biovector-