Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2014/12/15: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines</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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==Notes== | ==Notes== | ||
U2OS going into 3 vessels:<br> | |||
1. 1:10 passage into T-75 for cryo<br> | |||
2. 1:10 passage into T-75 for backup<br> | |||
3. 6-well plate (final conc. 2E5 cells/well, 4mL per well)<br><br> | |||
For the 1:10 passages, 9mL medium + 1mL cells<br><br> | |||
For the 6-well plate: | |||
* total volume = 4mL * (6 + 1 extra well) = 28 mL | |||
* Approx. 10E6 cells per T-75, or 1E6/mL | |||
* Desired final concentration of 2E5 cells/well, or 0.2mL of T-75 stock | |||
* 0.2*(6+1) = 1.4mL stock | |||
* in a 50mL conical, add 1.4mL of stock + 26.6mL of medium = 28mL total | |||
* aliquot 4mL diluted cells into each well, with 4mL left over | |||
<br><br> | |||
==Results/Conclusions== | ==Results/Conclusions== | ||
SKNSH culture is labeled PS-β and is in the upper incubator, middle shelf.<br><br> | SKNSH culture is labeled PS-β and is in the upper incubator, middle shelf.<br> | ||
U2OS flasks (2) and 6-well plate (1) are in the upper incubator, middle shelf.<br> | |||
U2OS cryo stocks will be prepped later in the week from the PS-1 flask.<br> | |||
<br> | |||
*'''[[User:David Benjamin Nyer|David Benjamin Nyer]] 18:18, 15 December 2014 (EST)''': | |||
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__NOTOC__ | __NOTOC__ |
Latest revision as of 00:36, 27 September 2017
RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines | Main project page Previous entry Next entry |
SummaryAfter starting the SKNSH cell lines in small (T25) flasks, they need to be transferred to a larger container to increase biomass in preparation of experimentation. If the initial thawing and starting of a cell line is Passage Alpha, the first transfer into a T75 flask is Passage Beta. The cells will need to have their medium removed, their attachment to the flask disrupted with trypsin-EDTA, and then be transferred into fresh media. Goals
Protocoladvancing adherent cells Materials
NotesU2OS going into 3 vessels: For the 1:10 passages, 9mL medium + 1mL cells For the 6-well plate:
Results/ConclusionsSKNSH culture is labeled PS-β and is in the upper incubator, middle shelf.
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