Difference between revisions of "PhiX:Serial dilution"
From CCGB
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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 | + | 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 | + | 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 | + | 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 #2 -> plate 100ul (1/10) -> 10<sup>-3</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 20:13, 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
