Head Binding of Tailspikes

CHP - March 13, 2000

Make heads c2ts(30) 9-am /13-am

1 Dilute heads to 4E9 phage equ /ml

2 Dilute tailspike 1/3 100 ul buffer DF 50 ul of tailspikes transfer 50ul to next tube,
do not forget to remove that 50ul from the last tube.

I started with 25 ug/ml, took 20ul into 180 ul (1/10 dilution) to give 200 ul of which

a) 50 ul for the head control
b) 50 ul to be passed to the next tube.

 

Therefore there is 100ul of tailspike 0.02ul x 25ug/ml x 0.2 /2. Divided by 2 because of the addition of heads serially diluted 10 times to 1.27E5 ug of tailspike.

Mix heads and tails 100ul and incubate 37oC about 1 hr.

Dilutions are:

Tube
Tspk concentration

Dilution for Plating
1
2.50E-01

6.1
2
8.33E-02

6.1
3
2.78E-02

6.1
4
9.26E-03

6.1
5
3.09E-03

6.5, 5.1
6
1.03E-03

4.5, 3.5
7
3.43E-04

3.5, 3.1
8
1.14E-04

2.1, 2.5
9
3.81E-05

1.5
10
1.27E-05

1.5

Add 2 ml of DF to each tube that had 200 ul ( tailspike + heads)
therefore, this is a 1/10 dilution for PFU at 30oC .