Tag Archives: PCR

NS5A-mCherry transfection with anti IgM stimulation, cells prepared for PCR

The WEHI 231 cells were transfected with NS5A-mCherry (100 uL). A control of Thapsigargin was used, cells in this group were treated with 87.5 (0.250mM) Thapsigargin for 4 hours before RNA isolation.

T=0 cells were collected an hour after stimulation. Cells were lysed with 500uL TRIzole reagent and frozen at -80C

Cells were stimulated with 35 uL of anti-IgM 12 hours after transfection.

T=36 cells were collected and lysed with 1 mL TRIzole reagent.

RNA from both the T=0 and T=36 samples was isolated according to TRIzole manufacturing protocol.

Nanodrop concentrations were as follows:

T=0 mock 225.5 ng/uL

T= mock stim 1956.5 ng/uL

T=0 NS5A 575 ng/uL

T=0 NS5A stim 1487 ng/uL

T=0 Thap 844.5 ng/uL

T=36 mock 24322 ng/uL

T=36 mock stim 400 ng/uL

T=36 NS5A 4415 ng/uL

T=36 NS5A stim 226 ng/uL

T=36 Thap 417.5 ng/uL

The integrity of the RNA was determined using a 1% agarose gel results to follow