Haynes Lab:Notebook/CRISPR Editing/2015/02/22: Difference between revisions

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Testing qPCR primers with dilution curves using Luc14 g023 treated rep 1 gDNA (82.4ng/ul)
Testing qPCR primers with dilution curves using Luc14 g023 treated rep 1 gDNA (82.4ng/ul)


{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''For'''
| align="center" style="background:#f0f0f0;"|'''Rev'''
| align="center" style="background:#f0f0f0;"|'''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''
'''Primer mix'''<br>
{| {{table}}
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Reagent'''
| align="center" style="background:#f0f0f0;"|'''Reagent'''
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|}
|}


''Template mixes''
'''Template mixes'''<br>
Make 1:10 dilution of gDNA sample so the new concentration is 8.24ul/ul.
Make 1:10 dilution of gDNA sample so the new concentration is 8.24ul/ul.
{| {{table}}
{| {{table}}

Revision as of 09:46, 23 February 2015

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02/22/2015

Testing qPCR primers with dilution curves using Luc14 g023 treated rep 1 gDNA (82.4ng/ul)

' 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
1:1 1:10 1:100 1:1000 1:10000 No temp ctrl
P set 1
P set 2
P set 3
P set 4
P set 5
P set 6


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