User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/24: Difference between revisions
From OpenWetWare
Line 23: | Line 23: | ||
* Add 2 ml/well fresh media with 1% Pen/strep | * Add 2 ml/well fresh media with 1% Pen/strep | ||
* Grow cells at 37°C (overnight) | * Grow cells at 37°C (overnight) | ||
Technical hint from Research Gate | |||
* In our lab, we concentrate our lentivirus after production using PEG-precipitation. This increases the concentration 50-100x. '''Then, we "spinoculate" K562 cells with a small amount of virus in an eppendorf tube. Typically, this means putting 50,000 - 100,000 K562 cells in an eppendorf tube, adding 1-2 uL of lentivirus, adding polybrene to 4 ug/ml, and centrifuging for 30 min at 800 x g (at 32 C or room temperature).''' If your lentivirus is of good quality, this method gives very high infection rate (normally I get greater than 90%...close to 100) | |||
** Mark Duncan, Northwestern U. | |||
Revision as of 07:13, 28 September 2014
Pc-TF Genomics | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
09/24/14
Transduction Day 2
|