Supplemental Data DEMETER DNA Glycosylase Establishes MEDEA Polycomb Gene Self-Imprinting by Allele-Specific Demethylation Mary Gehring, Jin Hoe Huh, Tzung-Fu Hsieh, Jon Penterman, Yeonhee Choi, John J. Harada, Robert B. Goldberg, and Robert L. Fischer Supplemental Experimental Procedures Primers for Bisulfite Sequencing Primers for the – 4 kb region were MEA3904 (5’-AACTTTATTCATRTAATRRTCRAACACT- 3’) or MEA3979 and MEA4510. The –3 kb region was amplified with MEA5187BFc (5’- CAAAATACTCTATTCTACATTCCCATCTAT-3’) and MEA5810BRc (5- TAAATAAATTAAATGAGTTTGAGTATAAAATG-3’), followed by a nested amplification with MEA5212 and MEA5810BRc. The –500 bp region was amplified with MEA7671 (5’- TAACCATTAAACATTAATTTAAATCTT-3’) or MEA7529 and MEA7935. MEA-ISR was amplified from Ler and Col-gl backgrounds using JP1026 and JP1027 (Cao and Jacobsen, 2002). A large deletion and extensive polymorphisms prevented the use of these primers in RLD. Instead, the first repeat was amplified with RLDBi (5’- TAATTTAAAATAATGGTGATGTTGTTAGTTTG-3’) and RLDBi4 (5’- AAAAARRTTTTATAAATATTAAATTAATATRA-3’). For MEA coding region bisulfite sequencing, Col-gl rosette leaf DNA was bisulfite treated as previously (Xiao et al., 2003) and methylation on the bottom strand determined. We sequenced 7 clones from MEA8355F (5’- TTTCACTCCAAACATATATAAATTAAC-3’) to MEA8755R (5’- GAYTAATGTATAAYTGTTTATTAGATGTAT-3’), 5 clones each from MEA8646F (5’- CTCTTCTRTATRTTTTTCTRAAAATTAARRA-3’) to MEA9066R (5’- TGYATYAATYTTGGYTTTTTTGGYTGAATG-3’) and from MEA9294F (5’- CACTTTTRTCRARAATRCAAAACCCACTT-3’) to MEA9801R (5’- TAATGYAAAAAYTAAYYATATAAATYGGTY-3’), 8 clones from MEA9810F (5’- CTTRATTATTAATTTRTARTCCATATTTAATAAACTR-3’) to MEA10221R (5’- GTGGYTAAATTAAAAAAGAAAGATTYAAAGTTAYYATG-3’), 10 clones from MEA10310F (5’-CCCRARTCTARATCCRTAARCATTAAATC-3’) to MEA10650R (5’- GGATYTGAGAYYAYAATYTTGTTTGATATAGAG-3’), 8 clones each from MEA10528F (5’-CTATTCCTTAATTACRTTTATTARTTACTRRT-3’) to MEA10905R (5’- GTTTTGTTAAGGTYTAATGAYATAGTAYATTG-3’) and MEA10761F (5’- TACTTACACTRTATTCCTTRATTATRC-3’) to MEA11285R (5’- TAYAAAYTYATGTTYAAATTAAATYTYATGG-3’), 6 clones from MEA11131F (5’- ATAARCACTACACACCATRCACTTRCAART-3) to MEA11460R (5’- CAAATTCTATAATCAAARTAATTCAAACC), 7 clones from MEA11571F (5’-