02/22/2015
Testing qPCR primers with dilution curves using Luc14 g023 treated rep 1 gDNA (82.4ng/ul)
'
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For
|
Rev
|
Target
|
Primer set 1 |
P207 |
P208 |
pX330
|
Primer set 2 |
P209 |
P210 |
pX330
|
Primer set 3 |
P211 |
P212 |
pX330
|
Primer set 4 |
P201 |
P202 |
GAPDH
|
Primer set 5 |
P203 |
P204 |
GAPDH
|
Primer set 6 |
P205 |
P206 |
MMP12
|
Primer mix
Reagent
|
Single well
|
x6.5
|
2x SYBR Green Master Mix |
7.5 |
48.75
|
750 nM F/R primer mix |
3 |
19.5
|
Total vol. |
10.5 |
68.25
|
Template mixes
Make 1:10 dilution of gDNA sample so the new concentration is 8.24ul/ul.
Dilution name
|
Make dilution stock
|
Make template MM
|
1:1 dil |
add 1.21ul to 8.79ul water |
6.5 ul dil to 22.75 H2O
|
1:10 dil |
add 1 ul to 9 ul water |
6.5 ul dil to 22.75 H2O
|
1:100 dil |
add 1 ul to 9 ul water |
6.5 ul dil to 22.75 H2O
|
1:1000 dil |
add 1 ul to 9 ul water |
6.5 ul dil to 22.75 H2O
|
1:10000 dil |
add 1 ul to 9 ul water |
6.5 ul dil to 22.75 H2O
|
Water only |
|
29.25 H2O
|
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1:1 |
1:10 |
1:100 |
1:1000 |
1:10000 |
No temp ctrl
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P set 1 |
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P set 2 |
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P set 3 |
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P set 4 |
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P set 5 |
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P set 6 |
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