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==Materials== {| {{table}} border=1 | align="center" style="background:#f0f0f0;"|'''Reagents and Kits''' | align="center" style="background:#f0f0f0;"|'''Source''' | align="center" style="background:#f0f0f0;"|'''Cat#''' |- | EZ DNA Methylation Gold Kit||Zymo Research||D5005 |- | 10x Ampligase buffer||Epicentre||A1905B |- | Ampligase||Epicentre||A3210K |- | dNTP||New England Biolab||N0447L |- | Stoffel fragment||Applied Biosystems||N808-0038 |- | ExoI||Epicentre||X40520K |- | ExoIII||Epicentre||EX4425K |- | 2xPhusion HF Master Mix||New England Biolab||F-531L |- | MmeI||New England Biolab||R0637L |- | Quick Ligation Kit||New England Biolab||M2200S |- | SYBR GreenI nucleic acid gel stain||Invitrogen||S7585 |- | Agencourt AMPure XP||Beckman Coulter||A63880 |- | 100% Ethanol || || |- | QIAQuick PCR Purification Kit || Qiagen || 28106 |- | E-Gel Size Select 2% || Invitrogen || G6610-02 |} {| {{table}} border=1 | align="center" style="background:#f0f0f0;"|'''Materials''' | align="center" style="background:#f0f0f0;"|'''Source''' | align="center" style="background:#f0f0f0;"|'''Cat#''' |- | Low Tube Strips, CLR||BioRad||TLS0801 |- | Microseal 'B' Film||BioRad||MSB1001 |- | Barrier Tips, or low retention pipette tips|| Neptune || BT200, BT10E, BT1000, BT20 |- | Mineral Oil|| Sigma|| M5904-500ml |- | 6% TBE Gel 1.0mmx10well ||Invitrogen|| EC6265BOX |- | 10x TBE buffer for PAGE || National Diagnostics || EC-860 |- | SYBR Gold nucleic acid gel stain||Invitrogen|| S11494 |} {| {{table}} border=1 | align="center" style="background:#f0f0f0;"|'''Primers''' | align="center" style="background:#f0f0f0;"|'''Sequence''' | align="center" style="background:#f0f0f0;"|'''Modifications''' |- | AmpF6.3NH2||/5AmMC6/CAGATGTTATCGAGGTCCGAC||5' Amino modifier C6 |- | AmpR6.3NH2||/5AmMC6/GGAACGATGAGCCTCCAAC||5' Amino modifier C6 |- | PE_t_N2, Y-adapter for MmeI ligation-top ||ACACTCTTTCCCTACACGACGCTCTTCCGATCTN*N||3'-Phosphorothioate bond |- | PE_b_A, Y-adapter for MmeI ligation-bottom ||/5Phos/AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAG || 5'-Phosphorylation |- | PCR_Fs|| AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTC || None |- | PCR_R.N2Ind1|| CAAGCAGAAGACGGCATACGAGATCGTGATCTCGGCATTCCTGCTGAACCGCTCTT || None |- | PCR_R.N2Ind2|| CAAGCAGAAGACGGCATACGAGATACATCGCTCGGCATTCCTGCTGAACCGCTCTT || None |- | PCR_R.N2Ind3|| CAAGCAGAAGACGGCATACGAGATGCCTAACTCGGCATTCCTGCTGAACCGCTCTT || None |- | PCR_R.N2Ind4|| CAAGCAGAAGACGGCATACGAGATTGGTCACTCGGCATTCCTGCTGAACCGCTCTT || None |}
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