Figure 3From: RNAi-mediated gene silencing in tick synganglia: A proof of concept studyIn vitro RNAi in tick synganglia. Adult female I. scapularis ticks were dissected and synganglia pooled in two groups and incubated with buffer alone or with (A) dsRNA-β-actin or (B) dsRNA Na+-K+-ATPase for 6 hrs at room temperature. Total RNA was extracted and cDNA produced. RT-PCR was conducted followed by gel electrophoresis. The I. scapularis Cyclophilin A or G was used as a normalizing factor (lanes 6–7). A) β-actin transcript level demonstrating the efficiency of RNAi in tick synganglia. (Lane 1, Low DNA Mass™ Ladder, 2–3: Mock and gene disrupted samples amplified with β-actin gene specific primers, 4–5: mock and gene silenced samples amplified with Na+-K+-ATPase gene specific primers, 6–7: mock and gene silenced amplified Cyclophilin A gene specific primers). B) Na+-K+-ATPase transcription demonstrated the efficiency of RNAi. (Lane 1, Low DNA Mass™ Ladder, 2–3: Mock and gene suppressed samples amplified with Na+-K+-ATPase gene specific primers, 4–5: mock and gene silenced samples amplified with β-actin gene specific primers, 6–7: mock and gene silenced samples amplified Cyclophilin G gene specific primers). Low DNA Mass™ ladder (100–2000 bp; 2000 bp, 1200 bp, 800 bp, 400 bp, 200 bp, 100 bp) was used from Invitrogen.Back to article page