SimpleChIP ® Human HSPA6 Promoter Primers were tested on DNA isolated from cross-linked cells using the SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. Real-time PCR was performed in duplicate on a serial dilution of 2% total input DNA (20 ng, 4 ng, 0.8 ng, and 0.16 ng) using a real-time PCR detection system and SYBR ® Green reaction mix. The PCR amplification efficiency (E) and correlation coefficient (R 2 ) were calculated based on the corresponding threshold cycle (C T ) of each dilution sample during 40 cycles of real-time PCR (95ºC denaturation for 15 sec, 65ºC anneal/extension for 60 sec).
PCR product melting curves were obtained for real-time PCR reactions performed using SimpleChIP ® Human HSPA6 Promoter Primers. Data is shown for both duplicate PCR reactions using 20 ng of total DNA. The melt curve consists of 80 melt cycles, starting at 55ºC with increments of 0.5ºC per cycle. Each peak is formed from the degradation of a single PCR product.