Haynes Lab:Notebook/Jan/2015/03/26: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==03/26/15== | ||
* Plasmid miniprep - 5 mL liquid culture for DPRE (D11.12) | * Plasmid miniprep - (2x) 5 mL liquid culture for DPRE (D11.12) | ||
* Assembly - BD006 into MV2 | * Assembly - BD006 into MV2 | ||
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---- | ---- | ||
'''Assembly''' | '''Assembly''' | ||
*'''[[User:Karmella Haynes|---Karmella]] 21:16, 26 March 2015 (EDT)''': | |||
* BD006/MV2: insert BD006(E/S)/4182 + vector MV2(E/X)/4529 | * BD006/MV2: insert BD006(E/S)/4182 + vector MV2(E/X)/4529 | ||
* Use Roche 2x buffer and NEB T4 ligase | * Use Roche 2x buffer and NEB T4 ligase | ||
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* Ligations | |||
# 10 uL rxn, vector + insert | # 10 uL rxn, vector + insert | ||
# 10 uL rxn, vector | # 10 uL rxn, vector | ||
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* 5 min./ ice | * 5 min./ ice | ||
* Plate on 100 ug/mL amp agar. Grow at 37 °C o/n | * Plate on 100 ug/mL amp agar. Grow at 37 °C o/n | ||
'''RESULTS''' (3/27/15) | |||
# BD006/MV2, 10 uL rxn, vector + insert: 6 colonies | |||
# BD006/MV2,10 uL rxn, vector: zero | |||
# BD006/MV2,15 uL rxn, vector + insert: 4 colonies | |||
# BD006/MV2,15 uL rxn, vector: zero | |||
* Success! Pick two colonies from each plate (1 and 3) for minipreps | |||
Latest revision as of 00:52, 27 September 2017
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03/26/15
Assembly
--> RT/ ~30 min.
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