Difference between revisions of "PhiX:Serial dilution"

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Line 4: Line 4:
 
Titer = x pfu/ml<br>
 
Titer = x pfu/ml<br>
 
OR =  y pfu in z ul
 
OR =  y pfu in z ul
 +
  
 
'''SERIAL DILUTION'''<br>
 
'''SERIAL DILUTION'''<br>
Add 100ul (10<sup>-1</sup>x) phage to 900ul HFB and mix = TUBE #1<br>
+
Add 100ul (10<sup>-1</sup>x) phage to 900ul HFB and mix -> tube #1<br>
Transfer 100ul (10<sup>-2</sup>x) #1 to 900ul HFB and mix = TUBE #2<br>
+
Transfer 100ul (10<sup>-2</sup>x) #1 to 900ul HFB and mix -> tube #2<br>
Transfer 100ul (10<sup>-3</sup>x) #2 to 900ul HFB and mix = TUBE #3<br>
+
Transfer 100ul (10<sup>-3</sup>x) #2 to 900ul HFB and mix -> tube #3<br>
 
and so on...
 
and so on...
 +
  
 
'''PLATING'''<br>
 
'''PLATING'''<br>
TUBE #1 -> plate 100ul (1/10) -> 10<sup>-2</sup>x<br>
+
Tube #1 -> plate 100ul (1/10) -> 10<sup>-2</sup>x<br>
TUBE #2 -> plate 100ul (1/10) -> 10<sup>-3</sup>x<br>
+
Tube #2 -> plate 100ul (1/10) -> 10<sup>-3</sup>x<br>
TUBE #3 -> plate 100ul (1/10) -> 10<sup>-4</sup>x<br>
+
Tube #3 -> plate 100ul (1/10) -> 10<sup>-4</sup>x<br>
 
and so on...
 
and so on...
 +
  
 
'''MATHS'''<br>
 
'''MATHS'''<br>

Revision as of 19:13, 6 August 2013

Summary

PHAGE SOLUTION
Titer = x pfu/ml
OR = y pfu in z ul


SERIAL DILUTION
Add 100ul (10-1x) phage to 900ul HFB and mix -> tube #1
Transfer 100ul (10-2x) #1 to 900ul HFB and mix -> tube #2
Transfer 100ul (10-3x) #2 to 900ul HFB and mix -> tube #3
and so on...


PLATING
Tube #1 -> plate 100ul (1/10) -> 10-2x
Tube #2 -> plate 100ul (1/10) -> 10-3x
Tube #3 -> plate 100ul (1/10) -> 10-4x
and so on...


MATHS
E.g. count n plaques on plate from 10-7x plate:
x = n * 10-7
OR, if z = 200ul (e.g., mutagenesis)
y = (n * 10-7) * z/1000