Alpha Diagnoestic Internationa

Strep/FLAG Tandem Affinity Purification (SF‐TAP) to Study Protein Interactions

. no. 2‐1000‐025) 1:10 in H 2 O (final concentration, 2 mM desthiobiotin) Anti–FLAG M2 agarose (Sigma‐Aldrich) FLAG elution

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

in 100µl of elution buffer at 65°C for 5 minutes;7.The eluted complexes can then be electrophoresed in denaturing PAGE and subjected to western blot analysis with an anti-Flag antibody for detection of the particular flag-tagged protein of choice (Figure

Genotyping using Affymetrix arrays

adaptor (5’-ATT ATG AGC ACG ACA GAC GCC TGA TCT-3’ and 5’phosphate AGC TAG ATC AGG CGT CTG TCG TGC TCA TAA-3’) (Affymetrix) are ligated to the digested DNAs with T4 DNA Ligase (New England BioLabs) in 25µL for 2 hours at 16°C. The ligations are stopped