Difference between revisions of "PhiX:Serial dilution"
From CCGB
| Line 3: | Line 3: | ||
'''PHAGE SOLUTION'''<br> | '''PHAGE SOLUTION'''<br> | ||
Titer = x pfu/ml<br> | Titer = x pfu/ml<br> | ||
| − | OR = y pfu in z ul | + | OR = y pfu in z ul<br> |
| − | |||
'''SERIAL DILUTION'''<br> | '''SERIAL DILUTION'''<br> | ||
| Line 10: | Line 9: | ||
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...<br> |
| − | |||
'''PLATING'''<br> | '''PLATING'''<br> | ||
| Line 17: | Line 15: | ||
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...<br> |
| − | |||
'''MATHS'''<br> | '''MATHS'''<br> | ||
Revision as of 20:14, 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
