Haynes Lab:Notebook/CRISPR Editing/2015/06/24: Difference between revisions

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(Autocreate 2015/06/24 Entry for Haynes_Lab:Notebook/CRISPR_Editing)
 
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==03//2015==
==06/24/2015==
 
'''Step 1: Diluting the DNA'''
 
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''ug of DNA for 3.2 wells'''
| align="center" style="background:#f0f0f0;"|'''ul plasmid'''
| align="center" style="background:#f0f0f0;"|'''water (32ul total)'''
|-
| g034 0.25||0.8||||
|-
| g034 0.5||1.6||||
|-
| g034 0.75||2.4||||
|-
| g034 1.0||3.2||||
|}
<br>
'''Step 2: make DNA + optimem mixes'''
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1 well'''
| align="center" style="background:#f0f0f0;"|'''3.2 x'''
|-
| g034 0.25||39||add 124.8ul optimem
|-
| g034 0.5||39||add 124.8ul optimem
|-
| g034 0.75||39||add 124.8ul optimem
|-
| g034 1.0||39||add 124.8ul optimem
|}
<br>
'''Step 3: add plus reagent to DNA+ opti'''
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1 well'''
| align="center" style="background:#f0f0f0;"|'''3.2 x'''
|-
| g034 0.25||1||add 3.2ul plus reagent
|-
| g034 0.5||1||add 3.2ul plus reagent
|-
| g034 0.75||1||add 3.2ul plus reagent
|-
| g034 1.0||1||add 3.2ul plus reagent
|}
<br>
'''Step 4: Make optimem + lipo mastermix'''
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1 well'''
| align="center" style="background:#f0f0f0;"|'''for 12 wells (x 13.2)'''
|-
| optimem||47||620.4
|-
| lipo||3||39.6
|}
<br>
'''Step 5: mix dna mix and lipo mix'''
{| {{table}}
| g034 0.25||add 160 lipo mix
|-
| g034 0.5||add 160 lipo mix
|-
| g034 0.75||add 160 lipo mix
|-
| g034 1.0||add 160 lipo mix
|}
'''Step 6: add 100ul to each well'''


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Revision as of 18:00, 24 June 2015

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06/24/2015

Step 1: Diluting the DNA

' ug of DNA for 3.2 wells ul plasmid water (32ul total)
g034 0.25 0.8
g034 0.5 1.6
g034 0.75 2.4
g034 1.0 3.2


Step 2: make DNA + optimem mixes

' 1 well 3.2 x
g034 0.25 39 add 124.8ul optimem
g034 0.5 39 add 124.8ul optimem
g034 0.75 39 add 124.8ul optimem
g034 1.0 39 add 124.8ul optimem


Step 3: add plus reagent to DNA+ opti

' 1 well 3.2 x
g034 0.25 1 add 3.2ul plus reagent
g034 0.5 1 add 3.2ul plus reagent
g034 0.75 1 add 3.2ul plus reagent
g034 1.0 1 add 3.2ul plus reagent


Step 4: Make optimem + lipo mastermix

' 1 well for 12 wells (x 13.2)
optimem 47 620.4
lipo 3 39.6


Step 5: mix dna mix and lipo mix

g034 0.25 add 160 lipo mix
g034 0.5 add 160 lipo mix
g034 0.75 add 160 lipo mix
g034 1.0 add 160 lipo mix

Step 6: add 100ul to each well


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