Haynes Lab:Notebook/CRISPR Editing/2015/08/27: Difference between revisions

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(Autocreate 2015/08/27 Entry for Haynes_Lab:Notebook/CRISPR_Editing)
 
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==03//2015==
==08/27/2015==
 
Set up annealing reaction for Surveyor of Luc14 dilution of g034 treated cells collected 7/8/15.
{|
| align="center" style="background:#f0f0f0;"|'''Cell type'''
| align="center" style="background:#f0f0f0;"|'''drug'''
| align="center" style="background:#f0f0f0;"|'''ug g034'''
| align="center" style="background:#f0f0f0;"|'''rep'''
| align="center" style="background:#f0f0f0;"|'''ul for 400ng'''
| align="center" style="background:#f0f0f0;"|'''ul buffer'''
| align="center" style="background:#f0f0f0;"|'''ul water for 15ul'''
|-
| Luc14||-||1||1||6.13||1.50||7.37
|-
| Luc14||-||1||2||6.03||1.50||7.47
|-
| Luc14||-||1||3||6.11||1.50||7.39
|-
| Luc14||-||0.1||1||5.98||1.50||7.52
|-
| Luc14||-||0.1||2||6.17||1.50||7.33
|-
| Luc14||-||0.1||3||6.60||1.50||6.90
|-
| Luc14||-||0.01||1||5.87||1.50||7.63
|-
| Luc14||-||0.01||2||6.08||1.50||7.42
|-
| Luc14||-||0.01||3||6.87||1.50||6.63
|}
<br>
Some PCR purifications from that same dilution experiment didn't have high concentrations so redid PCR with P163/215
:Gal4-EED + dox 0.1ul g034 rep 3
:Gal4-EED + dox 0.01ul g034 rep 1
<br><br>
Set up annealing reaction for Surveyor of Luc14 singlet from initial screen of gRNAs around the end of 2014
{|
| align="center" style="background:#f0f0f0;"|'''Luc14'''
| align="center" style="background:#f0f0f0;"|'''g029'''
| align="center" style="background:#f0f0f0;"|'''260'''
| align="center" style="background:#f0f0f0;"|'''280'''
| align="center" style="background:#f0f0f0;"|'''260/280'''
| align="center" style="background:#f0f0f0;"|'''ng/µL'''
| align="center" style="background:#f0f0f0;"|'''ul for 400ng'''
| align="center" style="background:#f0f0f0;"|'''ul ann buff'''
| align="center" style="background:#f0f0f0;"|'''ul H2O'''
|-
| Luc14||g030||0.096||0.052||1.855||95.552||4.19||1.5||9.31
|-
| Luc14||g033||0.049||0.027||1.802||48.747||8.21||1.5||5.29
|-
| Luc14||g036||0.057||0.03||1.9||56.608||7.07||1.5||6.43
|-
| Luc14||g040||0.067||0.035||1.927||66.681||6.00||1.5||7.50
|-
| Luc14||g040||0.054||0.029||1.848||53.968||7.41||1.5||6.09
|-
| Luc14||g041||0.072||0.038||1.864||71.533||5.59||1.5||7.91
|-
| Luc14||g042||0.042||0.023||1.865||42.202||9.48||1.5||4.02
|-
| Luc14||g043||0.082||0.045||1.793||81.554||4.90||1.5||8.60
|}




Revision as of 12:35, 27 August 2015

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08/27/2015

Set up annealing reaction for Surveyor of Luc14 dilution of g034 treated cells collected 7/8/15.

Cell type drug ug g034 rep ul for 400ng ul buffer ul water for 15ul
Luc14 - 1 1 6.13 1.50 7.37
Luc14 - 1 2 6.03 1.50 7.47
Luc14 - 1 3 6.11 1.50 7.39
Luc14 - 0.1 1 5.98 1.50 7.52
Luc14 - 0.1 2 6.17 1.50 7.33
Luc14 - 0.1 3 6.60 1.50 6.90
Luc14 - 0.01 1 5.87 1.50 7.63
Luc14 - 0.01 2 6.08 1.50 7.42
Luc14 - 0.01 3 6.87 1.50 6.63


Some PCR purifications from that same dilution experiment didn't have high concentrations so redid PCR with P163/215

Gal4-EED + dox 0.1ul g034 rep 3
Gal4-EED + dox 0.01ul g034 rep 1



Set up annealing reaction for Surveyor of Luc14 singlet from initial screen of gRNAs around the end of 2014

Luc14 g029 260 280 260/280 ng/µL ul for 400ng ul ann buff ul H2O
Luc14 g030 0.096 0.052 1.855 95.552 4.19 1.5 9.31
Luc14 g033 0.049 0.027 1.802 48.747 8.21 1.5 5.29
Luc14 g036 0.057 0.03 1.9 56.608 7.07 1.5 6.43
Luc14 g040 0.067 0.035 1.927 66.681 6.00 1.5 7.50
Luc14 g040 0.054 0.029 1.848 53.968 7.41 1.5 6.09
Luc14 g041 0.072 0.038 1.864 71.533 5.59 1.5 7.91
Luc14 g042 0.042 0.023 1.865 42.202 9.48 1.5 4.02
Luc14 g043 0.082 0.045 1.793 81.554 4.90 1.5 8.60



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