Iscript cdna reverse transcriptase kit

To determine if local expression of Fas and/or FasL was altered in response to HELLP syndrome mRNA was extracted from individual liver, placenta, kidney cortex, and kidney medulla as previously described [16 ]. Briefly, individual tissues (n=4–6/group) were crushed in liquid nitrogen, and mRNA was extracted using a Qiagen kit (Venlo, Netherlands) as outlined in the manufacturer’s instructions followed by treatment with DNAse per manufacturer’s directions (Applied Biosystems, Foster City, CA). cDNA was synthesized from 1μg of RNA with BioRad Iscript cDNA reverse transcriptase kit (Hercules, CA). Real-time polymerase chain reaction was performed using BioRad Sybre Green Supermix with an iCycler. Beta actin Ct (threshold cycle) was used as a housekeeping gene and the 2^−ΔΔCt method was used to analyze the results.
delta/delta Ct method was used to assess gene expression. Primer sequences [16 , 21 (link)] were provided by Life technologies (Carlsbad, CA).