User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/23: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</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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'''Transductions'''<br> | '''Transductions'''<br> | ||
Note: cells are in standard complete medium | |||
# Plate 1: U2OS | # Plate 1: U2OS | ||
# Plate 2: SK-N-SH | # Plate 2: SK-N-SH | ||
# Plate 4: K562 b (1 mL) - no one was around to consult on whether I should pellet the 2mL and replate | # Plate 4: K562 b (1 mL) - no one was around to consult on whether I should pellet the 2mL and replate | ||
Abbreviated protocol (from | Wells (for all plates) - added 400 μL virus | ||
* Add | # well 1 - GFP | ||
# well 2 - KAH160/015 (2010) | |||
# well 3 - KAH160/015 (2014) | |||
# well 4 - KAH165/015 (2010) | |||
# well 6 - KAH165/015 (2014) | |||
# well 7 - KAH170/015 (2014) | |||
Day 1: Abbreviated protocol (from Laura Gonzalez) | |||
* Retrieve frozen aliquots of harvested virus from -80C freezer (just outside TC room) | |||
* Thaw and quick-spin to get virus off of the inside of lid | |||
* Biohazard set-up: 10 mL Wescodyne waste conical (50 mL), 10 mL Wescodyne rinse | |||
* Remove 1mL of media from each well (except for K562) | |||
* Add 5ul of 2.2mg/ml Polybrene (thawed) to each well. Note: final concentration of Polybrene in well will be 8μg/ml after 400 μL viral aliquot is added | |||
* Spin at 2250 rpm for 25 minutes | |||
* Place the plates in the incubator (37°C) overnight. | |||
Latest revision as of 00:21, 27 September 2017
Pc-TF Genomics | Main project page Previous entry Next entry |
09/23/14
Transductions
Wells (for all plates) - added 400 μL virus
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