Haynes Lab:Notebook/CRISPR Editing/2015/03/24: Difference between revisions

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PCR purified


{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Cell type'''
| align="center" style="background:#f0f0f0;"|'''Treatment'''
| align="center" style="background:#f0f0f0;"|'''rep'''
| 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 200ng'''
| align="center" style="background:#f0f0f0;"|'''ul of hyb buff'''
| align="center" style="background:#f0f0f0;"|'''ul water'''
|-
| Luc14||Unt||1||0.037||0.019||1.922||36.604||5.46||1.50||8.04
|-
| Luc14||Unt||2||0.03||0.016||1.84||30.185||6.63||1.50||6.87
|-
| Luc14||Unt||3||0.028||0.015||1.845||27.725||7.21||1.50||6.29
|-
| Gal4-EED+dox||Unt||1||0.033||0.019||1.699||32.525||6.15||1.50||7.35
|-
| Gal4-EED+dox||Unt||2||0.032||0.019||1.674||31.888||6.27||1.50||7.23
|-
| Gal4-EED+dox||Unt||3||0.028||0.015||1.83||27.876||7.17||1.50||6.33
|-
| Luc14||g044||1||0.028||0.015||1.818||28.126||7.11||1.50||6.39
|-
| Luc14||g044||2||0.031||0.017||1.753||30.514||6.55||1.50||6.95
|-
| Luc14||g044||3||0.03||0.017||1.732||30.199||6.62||1.50||6.88
|-
| Gal4-EED+dox||g044||1||0.025||0.014||1.766||25.351||7.89||1.50||5.61
|-
| Gal4-EED+dox||g044||2||0.032||0.017||1.81||31.522||6.34||1.50||7.16
|-
| Gal4-EED+dox||g044||3||0.034||0.02||1.696||33.517||5.97||1.50||7.53
|-
| Luc14||g048||1||0.026||0.016||1.675||26.453||7.56||1.50||5.94
|-
| Luc14||g048||2||0.018||0.011||1.648||18.227||10.97||1.50||2.53
|-
| Luc14||g048||3||0.023||0.012||1.921||22.727||8.80||1.50||4.70
|-
| Gal4-EED+dox||g048||1||0.027||0.015||1.795||26.675||7.50||1.50||6.00
|-
| Gal4-EED+dox||g048||2||0.022||0.013||1.742||22.397||8.93||1.50||4.57
|-
| Gal4-EED+dox||g048||3||0.028||0.017||1.673||27.608||7.24||1.50||6.26
|}
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Revision as of 13:36, 24 March 2015

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03/24/2015

Ran on gel and did PCR purification kit for unt and g025 treated luc14 and gal4+dox cekks, nested PCR with P173/215.

Surveyor hybridization protocol on 200ng in 15ul

Cell type Treatment Rep 260 280 260/280 ng/µL ul for 200ng ul hyb buff ul water
Luc14 Unt 1 0.037 0.019 1.89 36.604 5.46 1.50 8.04
Luc14 Unt 2 0.025 0.014 1.745 25.137 7.96 1.50 5.54
Luc14 Unt 3 0.024 0.013 1.792 23.704 8.44 1.50 5.06
Gal4-EED+dox Unt 1 0.027 0.015 1.79 26.773 7.47 1.50 6.03
Gal4-EED+dox Unt 2 0.026 0.014 1.879 26.1 7.66 1.50 5.84
Gal4-EED+dox Unt 3 0.022 0.012 1.874 21.554 9.28 1.50 4.22
Luc14 g025 1 0.023 0.012 1.897 23.003 8.69 1.50 4.81
Luc14 g025 2 0.024 0.013 1.763 23.653 8.46 1.50 5.04
Luc14 g025 3 0.023 0.014 1.649 22.935 8.72 1.50 4.78
Gal4-EED+dox g025 1 0.023 0.013 1.76 23.111 8.65 1.50 4.85
Gal4-EED+dox g025 2 0.018 0.01 1.765 17.939 11.15 1.50 2.35
Gal4-EED+dox g025 3 0.026 0.016 1.59 26.08 7.67 1.50 5.83


Cut with 1ul Surveyor enzyme and 1ul enhancer. Ran on BA.


PCR purified

Cell type Treatment rep 260 280 260/280 ng/µL ul for 200ng ul of hyb buff ul water
Luc14 Unt 1 0.037 0.019 1.922 36.604 5.46 1.50 8.04
Luc14 Unt 2 0.03 0.016 1.84 30.185 6.63 1.50 6.87
Luc14 Unt 3 0.028 0.015 1.845 27.725 7.21 1.50 6.29
Gal4-EED+dox Unt 1 0.033 0.019 1.699 32.525 6.15 1.50 7.35
Gal4-EED+dox Unt 2 0.032 0.019 1.674 31.888 6.27 1.50 7.23
Gal4-EED+dox Unt 3 0.028 0.015 1.83 27.876 7.17 1.50 6.33
Luc14 g044 1 0.028 0.015 1.818 28.126 7.11 1.50 6.39
Luc14 g044 2 0.031 0.017 1.753 30.514 6.55 1.50 6.95
Luc14 g044 3 0.03 0.017 1.732 30.199 6.62 1.50 6.88
Gal4-EED+dox g044 1 0.025 0.014 1.766 25.351 7.89 1.50 5.61
Gal4-EED+dox g044 2 0.032 0.017 1.81 31.522 6.34 1.50 7.16
Gal4-EED+dox g044 3 0.034 0.02 1.696 33.517 5.97 1.50 7.53
Luc14 g048 1 0.026 0.016 1.675 26.453 7.56 1.50 5.94
Luc14 g048 2 0.018 0.011 1.648 18.227 10.97 1.50 2.53
Luc14 g048 3 0.023 0.012 1.921 22.727 8.80 1.50 4.70
Gal4-EED+dox g048 1 0.027 0.015 1.795 26.675 7.50 1.50 6.00
Gal4-EED+dox g048 2 0.022 0.013 1.742 22.397 8.93 1.50 4.57
Gal4-EED+dox g048 3 0.028 0.017 1.673 27.608 7.24 1.50 6.26


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