CUTANA pAG-Tn5是靶向剪切及轉(zhuǎn)座酶(CUT&Tag)技術(shù)中進(jìn)行高效繪制染色質(zhì)特征的關(guān)鍵試劑。與ChIP-seq相比，CUT&Tag在降低細(xì)胞需求量和測序深度的信噪比方面進(jìn)行了顯著改進(jìn)。CUTANA pAG-Tn5是一種高活性的E. coli轉(zhuǎn)座酶突變體(Tn5)與蛋白A/G的融合產(chǎn)物，可與來自多種物種宿主的靶抗體兼容。它也沒有表位標(biāo)簽，使其適合于標(biāo)簽介導(dǎo)的CUT&Tag(如FLAG、HA、TY1、V5等)。該產(chǎn)品經(jīng)過高度純化，已去除污染的E. coli?DNA，并在細(xì)胞數(shù)量少的情況下進(jìn)行復(fù)雜分析。為了每次實(shí)驗(yàn)的標(biāo)準(zhǔn)化以及抗體驗(yàn)證和反應(yīng)監(jiān)測，推薦使用SNAP-CUTANA??nucleosome spike-ins(如EpiCypher 19-1002)。Tn5帶有mosaic 接頭，可在CUT&Tag中使用。
Adapters
Tn5ME-A: 5’-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3’
Tn5ME-B: 5’-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3’
Tn5ME-rev: 5’-[phos]CTGTCTCTTATACACATCT-3’
組分
50 mM HEPES-KOH pH 7.2, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 50% glycerol.
保存條件
Stable for one year at -20℃?from date of receipt. The protein is not subject to freeze/thaw under these conditions.
數(shù)據(jù)示例
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FIGURE 1:? Protein gel data. CUTANA pAG-Tn5 for CUT&Tag (1 μg) was resolved via SDS-PAGE and stained ?with Coomassie blue. The migration and molecular ?weight of the protein standards are indicated. Uncharged ?pAG-Tn5 monomer is 78.5 kDa, however once charged ?with DNA, Tn5 dimerizes to a final complex weight of 191 kDa. |
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FIGURE 2: Size distribution of released chromatin. CUT&Tag was performed as described above. Recovered DNA was directly PCR amplified to produce sequenceready libraries. Agilent TapeStation traces for libraries derived from negative control IgG (top) and H3K27me3 (bottom) antibodies are shown. Excised DNA is highly enriched for mononucleosomes (peak at ~300 bp reflects ~150 bp insert size). |
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FIGURE 3:? CUT&Tag data. CUT&Tag was performed as described above. Representative sequencing tracks obtained using CUTANA pAG-Tn5 show a 229 kb close up view of the LAMC3 gene. CUTANA pAG-Tn5 produced clear peaks with genomic distribution profiles consistent with the known biological functions of H3K4me3 and H3K27me3 as well as minimal background in the IgG negative control. |
訂購詳情
貨號 | 產(chǎn)品名稱 | 規(guī)格 |
15-1017 | CUTANA? pAG-Tn5 for CUT&Tag | 50 Reactions |
15-1117 | CUTANA? pAG-Tn5 for CUT&Tag | 250 Reactions |
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