Daniel Clay Austin at 23:12, 15 January 2013

Daniel Clay Austin — Tue, 15 Jan 2013 23:12:44 GMT

←Older revision		Revision as of 23:12, 15 January 2013
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	Plating subclones for electrophysiological testing<br>		Plating subclones for electrophysiological testing<br>
	1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin and Zeocin) per colony picked.<br>		1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin and Zeocin) per colony picked.<br>
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	*Cell Media: Ham’s F12 from Gibco, +10% FBS, + 1% 100X Penicillin/Streptomycin		*Cell Media: Ham’s F12 from Gibco, +10% FBS, + 1% 100X Penicillin/Streptomycin
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Jon Sack: New page: Plating subclones for electrophysiological testing
1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin an...

Jon Sack — Tue, 09 Oct 2012 01:53:16 GMT

New page: Plating subclones for electrophysiological testing 1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin an...

New page

Plating subclones for electrophysiological testing 
1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin and Zeocin) per colony picked. 
2. Remove media from selected wells, rinse once with 200ul PBS, then add 50 ul trypsin. Watch to see when cells detach. 
3. While cells are trypsinizing, For each colony, put 1 ml media in a well of a 12-well plate, and put 2 ml media in a 35 mm tissue culture treated dish. Mark well and dish with name of a well a clone is to be picked from. 
4. Once they detach, add 200uL of selecting media. Triturate 5x to suspend cells. Remove 200ul suspension. Plate one drop of cell suspension in appropriately marked 35mm tissue culture dish. Place rest of cells in appropriate well of 12 well plate. 
5. Add 200 ul media to picked wells of 96-well plate 
6. Place cells incubator 

*Cell Media: Ham’s F12 from Gibco, +10% FBS, + 1% 100X Penicillin/Streptomycin

*Return to [[Sack:Protocols]]

Sack: Testing TREX Subclones Ephys - Revision history

Daniel Clay Austin at 23:12, 15 January 2013

Jon Sack: New page: Plating subclones for electrophysiological testing 1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin an...

Jon Sack: New page: Plating subclones for electrophysiological testing
1. Choose 12 wells on the 96-well plate that contain one colony. You will need 3.2 ml selecting media (F12, P/S, FBS, Blasticidin an...