Figure 2

Evaluation of granulocytes DNA extracted with separation medium after isolation of plasma and PBMCs. (A) A 0.8% agarose gel stained with ethidium bromide demonstrating high molecular weight DNA, and its near complete digestion with Eco RV and Bam H1. Lanes 1 and 24 represent the Lambda DNA/Hin dIII Marker. Lanes 2–21 are a side by side comparison of the digested and undigested granulocyte DNA extracted with separation medium from 10 subjects. Lanes 22–23 represent digested and undigested whole blood DNA for comparison with granulocyte DNA. (B) A typical performance of granulocyte DNA extracted with separation medium in real-time PCR for the 36B4 single copy gene (representative standard curve on the left and melting curves on the right); (C) A representative pyrogram for SNP rs6112 using granulocyte DNA extracted with separation medium for successful genotyping; (D) Gel-based VNTR analysis of the CCR5 VNTR on granulocyte DNA extracted with separation medium and whole blood DNA. Lanes 1 and 15 represent the 50–2,000 bp markers. Lanes 2–11 represent the granulocyte DNAs extracted with separation medium. Lane 12 represents the whole blood DNA and lanes 13 and 14 contain negative water controls. Arrows indicate expected product sizes (189 bp, and 157 bp) for this VNTR.