phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the presence of ATP
and Mg2+. The PIP3 product is detected by displacement of biotin-PIP3 from an energy transfer
complex consisting of Europium labelled anti-GST monoclonal
technologies, including hybridization probes
(e.g., FRET), hydrolysis probes (e.g., TaqMan®) and
displacement probes (e.g., molecular beacons).
• Initial denaturation for 20 sec at 95ºC is sufficient for
this source of contamination, a positive air
displacement hood that blows in filtered air over the workspace toward the investigator works well.
Separate solutions, pipettes, tubes, and tips should always
Equipment
The following equipment will be required for the trial:
• A peristatic pump/positive displacement pump
• One or two pressure gauges with Luer lock connection
• One roll of tubing (ideally
technologies, including hybridization probes (e.g.,
FRET), hydrolysis probes (e.g., TaqMan) and
displacement probes (e.g., molecular beacons).
• Use only gene-specific primers for one-step qRT-
PCR.
buffer rinse volume to be added into the final retentate.
To minimize dilution effects, buffer displacement/plug
flow with a completely full or drained system can also
be used to flush the product.