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[ for 18 hr . 1 Part 2 ( Determining the titre of bacteriophage ) |Label the five agar plates 102 10 106. N |Dispense 0. 9 ml of phage diluent (...

If the lysogenic plate is come from the dilution concentration of 10-6 solution and the total number of plaques count is 46, its total volume is 0.9ml, what is its titre of lysis within the patches of growth ?


what is the reason behind the plate for areas of lysis within patches of growth ?






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[ for 18 hr .
1
Part 2 ( Determining the titre of bacteriophage )
|Label the five agar plates 102 10 106.
N
|Dispense 0. 9 ml of phage diluent ( nutrient broth ) into each of the six sterile Eppendorf tubes ,
labelled 10 1 to 10% .
m
Make serial tenfold dilutions of the phage by dispensing 0.1 ml of the crude phage
preparation into the 10 1 tube , mix and transfer 0. 1 ml of 10 1 dilution to 10 2 tube . Each
transfer is to be done using a fresh tip and the process is repeated until 10% dilution is
reached .
Select a nutrient layer agar at 50 ' C , add 2 drops of S. epidermidis strain ( non -lysogenic strain )
using a Pasteur pipette . Then add 0. 1 ml of phage dilution 10% using a sterile tip ( the same
tip can be used for all phage dilutions ) . Mix the contents well and then pour the mixture on
to the surface of the corresponding agar plate ( 10% ) making sure that it is evenly distributed
by rocking the plate gently . The process is repeated with the other dilutions until the 102
dilution .
Allow all plates to solidify and then incubate at 37 ' C overnight .
in
MICZOLL
8

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dilated last week for evidence of lysis in or around the central
each patch of growth. Confirm which of the strains is lysogenic.
N
Select a patch which shows evidence of lysis, cut out the agar under the lysed area a
a sterile loop transfer the cut agar to a sterile centrifuge tube.
m
9 . Lml
Add 1 ml of nutrient broth to the centrifuge tube and use the end of the tip to br
agar in IL _
. tenth.
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