Haynes Lab:Notebook/CRISPR Editing/2015/02/03: Difference between revisions
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Rene M Davis (talk | contribs) (Autocreate 2015/02/03 Entry for Haynes_Lab:Notebook/CRISPR_Editing) |
Rene M Davis (talk | contribs) |
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==02//2015== | ==02/03/2015== | ||
Digested 400ng of purified PCR products from yesterday. Also duplicated with no enzyme, added 1ul elution solution: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Sample''' | |||
| align="center" style="background:#f0f0f0;"|'''Concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''ul for 400ng''' | |||
| align="center" style="background:#f0f0f0;"|'''ul Elut sol'n''' | |||
| align="center" style="background:#f0f0f0;"|'''ul buffer 4''' | |||
| align="center" style="background:#f0f0f0;"|'''ul PfoI''' | |||
|- | |||
| Luc14 g034 1||29.05||13.77||3.23||2||1 | |||
|- | |||
| Luc14 g034 2||28.85||13.87||3.13||2||1 | |||
|- | |||
| Luc14 g034 3||39.05||10.24||6.76||2||1 | |||
|- | |||
| GAL4-EED +dox g034 1||38.92||10.28||6.72||2||1 | |||
|- | |||
| GAL4-EED +dox g034 2||37.78||10.59||6.41||2||1 | |||
|- | |||
| GAL4-EED +dox g034 3||42.04||9.51||7.49||2||1 | |||
|} | |||
Let reaction go for 3 hours, diluted 1:10, ran on bioanalyzer | |||
<br> | |||
Hybridization for Surveyor, 400ng in 20ul reactions, 2 reactions each (for +/- nuc): | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Sample''' | |||
| align="center" style="background:#f0f0f0;"|'''Concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''ul for 400ng''' | |||
| align="center" style="background:#f0f0f0;"|'''ul elut sol'n''' | |||
| align="center" style="background:#f0f0f0;"|'''ul 10x hybridization buff''' | |||
|- | |||
| Luc14 g034 1||29.05||13.77||4.23||2 | |||
|- | |||
| Luc14 g034 2||28.85||13.87||4.13||2 | |||
|- | |||
| Luc14 g034 3||39.05||10.24||7.76||2 | |||
|- | |||
| GAL4-EED +dox g034 1||38.92||10.28||7.72||2 | |||
|- | |||
| GAL4-EED +dox g034 2||37.78||10.59||7.41||2 | |||
|- | |||
| GAL4-EED +dox g034 3||42.04||9.51||8.49||2 | |||
|} | |||
<br> | |||
Added 1ul surveyor nuclease, 1ul enhancer (made a MM). Added 2ul elution solution to -nuc samples. <br>Incubated at 42°C for 45 minutes.<br>Add 2ul stop solution to every tube (even -nuc).<br>make 1:10 dilution, analyze on bioanalyzer | |||
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|} | |} | ||
__NOTOC__ | __NOTOC__ |
Revision as of 09:04, 3 February 2015
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02/03/2015Digested 400ng of purified PCR products from yesterday. Also duplicated with no enzyme, added 1ul elution solution:
Let reaction go for 3 hours, diluted 1:10, ran on bioanalyzer
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