The reaction was carried out in 96-well optical reaction plates (Roche), using the Light Cycler 480 Sequence Detection System and software (Roche).
The reaction was carried out in 96-well optical reaction plates (Applied Biosystems, USA), using ABI Prism 7000 Sequence Detection System and software (PE Applied Biosystems, USA).
To obtain PCR conditions with reduced variability, premixes containing primers and probes and PCR master mix except template RNA were aliquoted into 96-well optical reaction plates.
Relative quantification using standard curve method was performed for each primer pair and 96-well Optical Reaction plates were read in an Applied Biosystems AbiPrism 7300 real-time PCR instrument (Absolute Quantification Method).
Quantitative real-time PCR reactions were performed for each primer pair and 96-well Optical Reaction plates were read in an Applied Biosystems StepOnePlus Real-Time PCR System (Comparative Ct (ΔΔCt) Quantification Method).
Five µl (approximately 30 ng) of cDNA and 20 µl of the master mix solution were added to each well of a 96 well optical reaction plate (Applied Biosystems).
Real-time reactions were performed in 96-well optical reaction plates (SABiosciences) by adding RT2 SYBR Green/ROX (SABiosciences) and sterile ddH2O to each reaction that had a final volume of 25 µ l.
A one step real time PCR using the KCNMA1 primers (Qiagen) was performed in special 96-well TaqMan optical reaction plate format on a ABI Prism 7000 sequence detection system (Applied Biosystems, Foster city, CA, USA).
Relative quantification using the standard curve method was performed for each primer pair and 96-well Optical Reaction plates were read in an Applied Biosystems AbiPrism 7300 real-time PCR instrument (Absolute Quantification Method).
20 µl reactions were mixed in 96-well ABI Optical Reaction Plates as follows: 10 µl 2× Reaction Mix, 2 µl forward primer and 2 µl reverse primer (100 nM final), 0.16 µl RT mix, 1 µl RNA (70 ng total) and 4.84 µl H2O.
Every reaction was run in triplicate in a 384-well clear optical reaction plate.
