赛默飞世尔Thermo Fisher SIGN'DNGR L

Combined 3C-ChIP-Cloning (6C) Assay: A Tool to Unravel Protein-Mediated Genome Architecture

analysis. Restriction enzyme buffer (10X) Dilute the 10X stock to 1.14X for Steps 12 and 13. RNase A, DNase-free (10 mg/mL; Sigma R6513) SDS (Sodium dodecyl sulfate; 20% w/v; Fisher BP166) Sodium acetate (3 M, pH 5.2

Single Primer ("Semi-Random") P

to about 1 ng/µL 3 µL "T/TS" @ 0.4 u Taq/µL -------- 30 µL Cycle as follows: 30"x94°C 20 cycles 0"x94°C 0"x55°C 1'x72° S=9 30 cycles 0"x94°C 0"x40°C 1'x72° S=6 30 cycles 0"x94°C 0"x55°C 1'x72° S=9 Clean up: Add 1 µL ExoI

RNA-biotin based pulldown assays for the detection of siRNA targeted genomic regions and siRNA directed histone modifications (PROT32)

upon the dual pulldown to incorporate a third pulldown which is an iteration of the ChIP and is a pulldown for H3K27me3+ (Figure 1b). The third assay described here is the biotin-RNA pulldown of a low-copy RNA that spans the siRNA targeted promoter region