User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/09: Difference between revisions
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My ORF-carrying plasmids: | My ORF-carrying plasmids (& 6-well plate labels): | ||
# KAH160/015 (8/09/10) | # KAH160/015 (8/09/10) | ||
# KAH160/015 (9/07/14) --> new miniprep | # KAH160/015 (9/07/14) --> new miniprep | ||
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# KAH170/015 (8/09/10) | # KAH170/015 (8/09/10) | ||
# KAH170/015 (9/07/14) --> new miniprep | # KAH170/015 (9/07/14) --> new miniprep | ||
# Turbo GFP control, 980 ng/μL, used 2.5 μL (LaBaer lab) | |||
* New minipreps had low concentrations (~20 ng/μL, 100 μL total) | * New minipreps had low concentrations (~20 ng/μL, 100 μL total) | ||
** Zymo-cleaned total minipreps (9/07/14 samples) to get ~2000 ng | ** Zymo-cleaned total minipreps (9/07/14 samples) to get ~2000 ng in 14 μL H<sub>2</sub>O | ||
** Also measured conc. of old plasmids (8/09/10) - was in a hurry, so no time to clean and concentrate old plasmids | ** Also measured conc. of old plasmids (8/09/10) - was in a hurry, so no time to clean and concentrate old plasmids | ||
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| KAH160/015 8/09/10 || 1.210 || 1.89 || 210.2 || 2522.4/ 12 μL + 2 μL H<sub>2</sub>O | | KAH160/015 8/09/10 || 1.210 || 1.89 || 210.2 || 2522.4/ 12 μL + 2 μL H<sub>2</sub>O | ||
|} | |} | ||
Lipo complexes (done in LaBaer TC room) | |||
* 10 μL of ORF/dR8.91/VSVG mix | |||
* + 10 μL DEPC water | |||
* + 570 μL OptiMEM (Haynes Lab) | |||
* + 2.5 μL PLUS reagent (Haynes Lab) --> 5 min. room temp | |||
* + 7.5 μL Lipo LTX (Haynes Lab) --> 30 min. room temp | |||
Revision as of 17:00, 9 September 2014
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09/09/14
Lenti Transfections
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