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钠洪
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3 真心說用過 Zymo Kits 畢業以後才不會後悔 TM EZ DNA Methylation Kits EZ DNA MethylationTM selection guide bisulﬁte treatment 03 效率最高 DNA 回收率最高 DNA Methylation - Bisulﬁte Methods DNA Input 100 pg - 2 µg (optimal input : ng) 高達 99% CT conversion 效率利用 spin column 純化 CT conversion DNA 回收後產物適用 PCR sequecing microarray 等彈性選擇可使用 Gold Direct 或 Lightning 系統大幅縮短操作時間

4 DNA Methylation - Bisulfite Methods Zymo Low Elution Desulphonation Column Zymo bisulfite DNA conversion kit Zymo-Spin TM IC column, column desulphonation, bisulfite-treated DNA 5 µg DNA, 6 µl,, buffer, DNA bisulfite conversion, non-methylated cytosine uracil; DNA PCR,uracil thymine, CT conversion 5000 Illumina Sequenom Zymo Kit CT conversion DNA EZ DNA Methylation TM Kit DNA > 99% > 80% hours D rxns D rxns EZ DNA Methylation-Gold TM Kit DNA > 99% > 75% 3 hours D rxns D rxns EZ DNA Methlyation-Lightning TM Kit DNA > 99.5% > 80% 1.5 hours D rxns D rxns EZ DNA Methlyation-Direct TM Kit DNA, cells, blood, D rxns > 99.5% > 80% 4 hours tissue, FFPE D rxns 04

5 Zymo Kit! Pico Methyl-Seq TM Library Prep Kit Post-Bisulfite Library Preparation WGBS (Whole Genome Bisulfite Sequencing) (post-bisulfite library) DNA input FFPE : ligation, 6 : (bisulfite conversion enzymes index barcodes purification columns) Specifications DNA Input...10 pg-100 ng DNA Samples...genomic DNA, FFPE DNA Sequencing Platform Compatibility:...Illumina's TruSeq chemistries for HiSeq TM and MiSeq TM sequencing platforms DNA Methylation - Bisulfite Methods Agilent 2200 TapeStation D1K libraries (A) Electropherogram of a Pico Methyl-Seq (WGBS) library prepared using 100 pg of human gdna. (B) Gel of libraries prepared from varied amounts of human gdna. Fragments ranged between 150 and 350 bp. Citation Schroeder DI et al. (2015). Early Developmental and Evolutionary Origins of Gene Body DNA Methylation Patterns in Mammalian Placentas. PLoS Genet. Aug 4;11(8). D preps Pico Methyl-Seq TM Library Prep Kit D preps 05

CpG:MSREs dsdna,qpcr Ct reference qpcr DNA, bisulfite conversion One-tube, :MSREs digestion qpcr single-, multi-locus DNA methylation human methylated/non-methylated DNA standard control primers,

6 DNA Methylation - Non-Bisulfite Methods Test Reaction Reference Reaction!! OneStep TM qmethyl TM Kits Region Specific 5-mC Analysis MSREs (Methylation Sensitive Restriction Enzymes), CpG non-methylated CpG:MSREs dsdna,qpcr Ct reference ( MSREs degestion ) methylated CpG:MSREs dsdna,qpcr Ct reference qpcr DNA, bisulfite conversion One-tube, :MSREs digestion qpcr single-, multi-locus DNA methylation human methylated/non-methylated DNA standard control primers, Specifications Format Well Plate DNA Input...20 ng in 5 µl Processing Time...~4 hours Add DNA Real-time PCR Simple, One Step 4 hr, DNA Non-methylated DNA Input Methylated DNA Input 1 non-methylayed methylayed DNA sample 2 DNA MSREs digestion, test reaction; DNA MSREs digestion, reference reaction 3 primer set, qpcr ( SYBR green Taqman probe ) 4 non-methylated DNA: Ct Test > Ct Ref methylated DNA: Ct Test Ct Ref Citation van Dijk M et al. (2012) Naturally occurring variation in trophoblast invasion as a source of novel (epigenetic) biomarkers. Front Genet.; 3:22. OneStep qmethy TM (with SYTO 9 dye for qpcr) D x 96 well OneStep qmethyl TM -Lite (without SYTO 9 dye) D x 96 well 06

7 Zymo Kits, 5-mC TM DNA ELISA Kit Global 5-mC Quantification : ELISA 5-mC, : purified genomic DNA,PCR,DNA plasmid DNA : 0.5% 5-mC per 100 ng input DNA Specifications DNA Input ng Detection % 5-mC per 100 ng Assay Time hr DNA Methylation - Antibody Methods 5-mC DNA ELISA DNA 5-mC, mass spectrometry (MS) Citations Chiara Riganti et al. (2013). Temozolomide downregulates P-glycoprotein expression in glioblastoma stem cells by interfering with the Wnt3a/glycogen synthase-3 kinase/b-catenin pathway. Neuro-Oncology 15(11): Alexandra Paradis et al. (2014). Endothelin-1 promotes cardiomyocyte terminal differentiation in the developing heart via heightened DNA methylation. Int J Med Sci.11(4): D5325 1x96-well 5-mC TM DNA ELISA Kit D5326 2x96-well 07

8 DNA Methylation - Antibody Methods ~ Zymo Kit Methylated-DNA IP Kit Global 5-mC Quantification anti-5-methylcytosine monoclonal antibody enrich DNA control DNA primers, DNA PCR whole-genome amplification ultra-deep sequencing microarray Specifications Format...Magnetic Beads Optimal DNA Input ng Elution Volume...10 µl Enrichment Factor...> 100 fold Processing Time...4 hr 08

Citations Sunahori, K., et al. (2013) PP2Ac promotes DNA hypomethylation by suppressing the pmek/perk/dnmt1 pathway in T cells from controls and SLE patients. J. Biol. Chem. Eichten SR et al.

9 Citations Sunahori, K., et al. (2013) PP2Ac promotes DNA hypomethylation by suppressing the pmek/perk/dnmt1 pathway in T cells from controls and SLE patients. J. Biol. Chem. Eichten SR et al. (2012) Spreading of heterochromatin is limited to specific families of maize retrotransposons. PLoS Genet. 8(12). enrich DNA DNA Methylation - Antibody Methods Salmon sperm genomic DNA "spiked" with non-methylated/methylated control DNA was processed with the kit and the eluted DNA was amplified by PCR using the supplied control primers. Digestion of the amplicons with Nco I produced two 175 bp fragments for the methylated DNA control or one 350 bp fragment for the nonmethylated DNA control. Methylated-DNA IP Kit D preps Taq polymerase enriched DNA? "hot start" polymerase, ZymoTaq DNA Polymerase, cycles, ZymoTaq PCR Methylated DNA was immunoprecipitated using the Methylated-DNA IP Kit. The resultant DNA was used in a PCR assay comparing Zymo Research's hot start ZymoTaq TM polymerase vs. that of three other suppliers. Expected amplicon size is 350 bp. 09

10 RNA Methylation - Bisulfite Methods,, Kit EZ RNA Methylation TM Kit RNA bisulfite conversion, RNA non-methylated cytosine! RNA input Converted RNA RT-PCR sequencing library preparation Next-Gen sequencing RNA Methylation? RNA methylation RNA, epigenetic,, RNA, RNA, 5-methylcytosine in RNA,Zymo Research EZ RNA Methylation Kit, RNA methylation Original RNA with Methylated CpG : Converted RNA : cdna Sequence : EZ RNA Methylation Kit, RNA bisulfite conversion, RNA RT-PCR,clone 7 methylated cytosine bisulfite conversion, cytosine, 3, 4, 13 non-methylated cytosine uracil, RT-PCR,uracil thymine In-column Desulphonation Input RNA Denature RNA Bisulfite Conversion For RNA purification, use Direct-zol RNA Miniprep or Quick-RNA Miniprep Converts non-methylated cytosine into uracil. Methylated cytosine remains unchanged during the treatment. Elute Bisulfite-treated RNA Ready for Analysis Ideal for RT-PCR, sequencing, library preparation and Next- Gen sequencing Citations Amort T, Soulière MF, Wille A, Jia XY, Fiegl H, Wörle H, Micura R, & Lusser A (2013). Long non-coding RNAs as targets for cytosine methylation. RNA biology, 10 (6). Schosserer, M. et al. (2015) Methylation of ribosomal RNA by NSUN5 is a conserved mechanism modulating organismal lifespan. Nat Commun. 6, Product Cat No Size R5001 EZ RNA Methylation Kit 50 preps R5002 EZ RNA Methylation Kit 200 preps 10

12 我的甲基化時代 Methylation Time 本 DM 產品至止台北折 ANTEC Bioscience Inc 新竹台中台南高雄 U/A/UV040922

TRUSTED BRAND Infinium and GoldenGate assay systems SureSelect Methyl-seq Target Enrichment System MassARRAY EpiTYPER NimbleGen SeqCap Epi Enrichment

TRUSTED BRAND Infinium and GoldenGate assay systems SureSelect Methyl-seq Target Enrichment System MassARRAY EpiTYPER NimbleGen SeqCap Epi Enrichment We are TWINS!! Epigenetics Secret Garden Why are we so different? TEL (02)2720-2215 TEL (05)2844-162 TEL (03)6684-586 TEL (06)2890-665 TEL (04)2463-3591 TEL (07)3470-143 免付費客服專線 TRUSTED BRAND Infinium