Name: genome-wide determination of mammalian replication timing by dna content measurement
Uploaded: 2017-01-19T14:00:00
Description: Watch this Scientific Journal Video about Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement at JoVE.com

私たちは、数字を生成する際に支援するためのオリヤー語ヴァルディに感謝します。 ISグループでの作業は、イスラエル科学財団（助成番号10分の567）とグラント（＃281306）を開始し、欧州研究評議会によってサポートされていました。

注：Torは唯一の成長、同期されていない細胞で測定することができます。通常、S期にある〜1×10 5個の細胞になり、2×10 6急成長する細胞、（律速段階） -手順は、少なくとも1で開始する必要があります。 2または3つの複製を使用して、各実験を実施することをお勧めします。 CNR-のToRの全体のプロセスは、1週間以内に完了することができます - 2日には、ライブラリ?...

<ol>
	<li>Farkash-Amar, S., Simon, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+analysis+of+the+replication+program+in+mammals.">Genome-wide analysis of the replication program in mammals.</a> Chromosome Res. 18 (1), 115-125 (2010).</li><li>Yaffe, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+analysis+of+DNA+replication+timing+reveals+conserved+large-scale+chromosomal+architecture.">Comparative analysis of DNA replication timing reveals conserved large-scale chromosomal architecture.</a> PLoS Genet. 6 (7), e1001011 (2010).</li><li>Hiratani, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+reorganization+of+replication+domains+during+embryonic+stem+cell+differentiation.">Global reorganization of replication domains during embryonic stem cell differentiation.</a> PLoS Biol. 6 (10), (2008).</li><li>Rivera-Mulia, J. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenomic+replication:+linking+epigenetics+to+DNA+replication.">Epigenomic replication: linking epigenetics to DNA replication.</a> Bioessays. 25 (7), 647-656 (2003).</li><li>Sima, J., Gilbert, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complex+correlations:+replication+timing+and+mutational+landscapes+during+cancer+and+genome+evolution.">Complex correlations: replication timing and mutational landscapes during cancer and genome evolution.</a> Curr Opin Genet Dev. 25, 93-100 (2014).</li><li>Kenigsberg, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mutation+spectrum+in+genomic+late+replication+domains+shapes+mammalian+GC+content.">The mutation spectrum in genomic late replication domains shapes mammalian GC content.</a> Nucleic Acids Res. 44 (9), 4222-4232 (2016).</li><li>Woo, Y. H., Li, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+replication+timing+and+selection+shape+the+landscape+of+nucleotide+variation+in+cancer+genomes.">DNA replication timing and selection shape the landscape of nucleotide variation in cancer genomes.</a> Nat Commun. 3, 1004 (2012).</li><li>Liu, L., De, S., Michor, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+replication+timing+and+higher-order+nuclear+organization+determine+single-nucleotide+substitution+patterns+in+cancer+genomes.">DNA replication timing and higher-order nuclear organization determine single-nucleotide substitution patterns in cancer genomes.</a> Nat Commun. 4, 1502 (2013).</li><li>Goren, A., Cedar, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replicating+by+the+clock.">Replicating by the clock.</a> Nat Rev Mol Cell Biol. 4 (1), 25-32 (2003).</li><li>Selig, S., Okumura, K., Ward, D. C., Cedar, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delineation+of+DNA+replication+time+zones+by+fluorescence+in+situ+hybridization.">Delineation of DNA replication time zones by fluorescence in situ hybridization.</a> EMBO J. 11 (3), 1217-1225 (1992).</li><li>Smith, L., Thayer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chromosome+replicating+timing+combined+with+fluorescent+in+situ+hybridization.">Chromosome replicating timing combined with fluorescent in situ hybridization.</a> J Vis Exp. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coating+agents+for+cell+recovery.">Coating agents for cell recovery.</a> Google Patents. , (1993).</li><li>Koren, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+relationship+of+DNA+replication+timing+to+different+forms+of+human+mutation+and+variation.">Differential relationship of DNA replication timing to different forms of human mutation and variation.</a> Am J Hum Genet. 91 (6), 1033-1040 (2012).</li><li>Rhind, N., Gilbert, D. 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CNR-Torは（Rhind N.およびGilbert DM 20によってレビュー）SにFACSおよびG1相により分割することができる任意の真核生物増殖細胞集団の原理で行うことができます。ここに記載される方法は、ヒト及びマウスのような約3ギガビットのゲノムサイズを有する哺乳動物細胞に調整しました。 （細胞調製および配列決定の深さ）CNR-のToRプロトコルにおける小さな変化は、他の真核生物?...

哺乳動物のDNA複製は、細胞周期の間に正確に一度、各染色体の正確な複製を確保するために厳密に調節されています。レプリケーションは、高度に制御順序に従って発生-他のゲノム領域は、中間または後期S期（中期および後期複製ドメイン）で、後に複製する一方、複数の大規模なゲノム領域（〜Mbが）、S期（早期複製ドメイン）の先頭に複製する1。ゲノムの50％、また、癌の形質転換5中分化3,4の間及びより少ない程度に、組織2との間ののToRを変更-ゲノムのほとんど30％が、全ての組織（構成のToRドメイン）で同時に複製します。また、特定のゲノム領域は非同期6、7、8、すなわち差が複製します2対立遺伝子間のToRインチ Torは転写レベル、GC含量、クロマチン状態、遺伝子密度など 1,9を含む多くのゲノムおよびエピ機能と相関します。 ToRのも突然変異率とタイプ10、11、したがって、当然に関連付けられている、複製プログラムの摂動は、がん12、13にリンクされています。 ToRとクロマチン構造の間の因果関係は未だ解明されていません。開いたクロマチンが早い複製を容易にすることが可能です。しかし、代替モデルは14、クロマチンは、初期および後期複製領域1のパッケージングを差動にS期リードの開始時と終了時に複製し、異なるクロマチン制御因子存在の間に組み立てられていることを示唆しています</sup&gt;。我々は最近のToRは異なるゲノム領域11に発生する突然変異のタイプに影響を与えることによって、GC含量を整形することが示されています。 蛍光in situハイブリダイゼーション（FISH）は、個々の遺伝子座でのToRを測定するための主要な方法です。これは、単一のFISHシグナル対与えられた対立遺伝子15、16二重の割合を示すS期の細胞の割合をカウントすることによって簡単に行われます。別の方法は、Sに沿って複数の時点にそれらのDNA含有量に応じて細胞を選別、BrdUでDNAを標識のBrdUを含むDNAを免疫沈降し、定量PCR 17で沈殿したDNAの存在量をチェックするパルスで構成されています。 ゲノムのToRマッピングは、2つの方法によって達成することができます。第一の方法は、上述したBrdU-IPに基づく方法のゲノムのバージョンであり、ここで量の定量各分画中の沈殿したDNAは、マイクロアレイへのハイブリダイゼーションを介して、全ゲノムまたはディープシークエンシングにより同時に行われます。第二の方法、CNR-Torは、G1細胞におけるDNA含有量によってS期細胞と正規化の各ゲノム領域のコピー数を測定することに基づいています。この方法では、細胞は非複製（G1期）および（S期）基（ 図1）の複製にFACSによってソートされます。 G1の細胞はすべてのゲノム領域において同じコピー数を持っているので、それらのDNA含有量は同じである必要があります。一方、SにおけるDNAコピー数が遅く複製領域は、したがって、それらのDNA含量がするほとんどの細胞ではまだ複製していない一方で、初期の複製領域は、ほとんどの細胞で複製を受け、したがって、それらのDNA含有量が倍になるので、のToRに依存しますG1細胞のものと同様です。したがって、DNA含量のG1比Sは、のToRを示します。各ゲノム領域のためのDNAの量は、ハイブリダイゼーションによってのいずれかで測定されますマイクロアレイまたはディープシークエンシング2、8による。 CNR-のToRの方法の利点をさらに説明します。 図2で説明したように本論文では、ゲノムのToRマッピング用のCNR-のToR方法について説明します。紙は、結果の基本的な分析およびゲノムのToRマップの作成まで、細胞を採取からプロセス全体の細部を説明します。この論文に記載されているプロトコルが正常に培養で増殖させた種々の細胞型上で実行されています。このプロトコルの将来の改善は、in vivoでのToRのマッピングに、希少細胞タイプにつながることができます。

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			<td height="21" style="height:21px;width:339px;">PBS</td>
			<td style="width:157px;">BI (Biological Industries)</td>
			<td style="width:119px;">02-023-1A</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypsin-EDTA</td>
			<td>BI (Biological Industries)</td>
			<td>03-052-1B</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15ml conical tube</td>
			<td>Corning</td>
			<td align="right">430790</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5ml Polystyrene round Bottom tube with cell strainer cap&#160;</td>
			<td>BD-Falcon</td>
			<td align="right">352235</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol</td>
			<td>Gadot</td>
			<td>64-17-5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RNAse-A 10mg/ml</td>
			<td>Sigma</td>
			<td>R4875</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Propidiom iodide 1mg/ml</td>
			<td>Sigma</td>
			<td>P4170</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">parafilm</td>
			<td>Parafilm</td>
			<td>PM-996</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1.5ml DNA LoBind Eppendorf tubes&#160;</td>
			<td>Eppendorf</td>
			<td align="right">22431021</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BSA</td>
			<td>Sigma</td>
			<td>A7906</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1.7ml MaxyClear tube&#160;</td>
			<td>Axygen</td>
			<td>MCT-175-C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">magnetic beads - Agencourt AMPure XP&#160;</td>
			<td>Beckman Coulter</td>
			<td>A63881</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ultrasonicator</td>
			<td>Covaris</td>
			<td>M-series&#160; -530092</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">50 &#181;l microTUBE AFA Fiber Screw-Cap 6x16mm</td>
			<td>Covaris</td>
			<td align="right">520096</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Qubit fluorometer</td>
			<td>Invitrogen</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Qubit dsDNA High Sensitivity (HS) Assay Kit</td>
			<td>Invitrogen</td>
			<td>Q32854</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Electrophoresis.2200 Tape station system</td>
			<td>Agilent</td>
			<td>D1000 ScreenTape</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Seqeuncing - Illumina NextSeq system</td>
			<td>Illumina</td>
			<td>SY-415-1001</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dneasy kit for DNA purification</td>
			<td>Qiagen</td>
			<td align="right">69504</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PureProteom Magnetic Stand</td>
			<td>Millipore</td>
			<td>LSKMAGS08</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anti-BrdU/FITC</td>
			<td>DAKO</td>
			<td>F7210</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FACS sorter</td>
			<td>BD</td>
			<td>FACSARIA III</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FACS software</td>
			<td>BD</td>
			<td>FACSDiva v 8.0.1</td>
		</tr>
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genome-wide determination of mammalian replication timing by dna content measurement

ゲノムの複製は、DNA複製の忠実度を保証する高度に調節されたプロセスにおける細胞周期のS期の間に起こります。それぞれのゲノム領域は、複製の複数の起源の同時活性化を介して、S期の間に明確な時に複製されます。レプリケーション（ToRの）の時間は、多くのゲノムおよびエピジェネティックな機能と相関し、突然変異率やがんにリンクされています。健康と病気で、複製プログラムの完全なゲノムビューを理解することは、主要な将来の目標や課題です。 、哺乳動物細胞のゲノムワイドのToRをマップするための簡単な方法：この記事では詳細に（CNR-のToRここでいう）メソッド」レプリケーションのゲノム時間をマッピングするためのS / G1のコピー数の比率」を説明しています。この方法は、S期細胞及びG1期の細胞の間のコピー数の差異に基づいています。 CNR-のToR法は6工程で行われる：ヨウ化プロピジウム（PI）で細胞を染色の調製。 2.ソル（FACS）を蛍光活性化細胞選別を用いた定G1及びS期細胞。 3. DNA精製。 4.超音波処理; 5.ライブラリの準備および配列決定;そして6.バイオインフォマティクス解析。 CNR-のToR方法は、詳細なレプリケーション・マップになり、迅速かつ簡単なアプローチです。

典型的なToRのマップは、マウス胚性線維芽細胞（MEF）は、 図3に示されています。それは個々のウィンドウのための正規化されたS / G1比（ステップ8.3）、ならびに立方平滑化及び補間（ステップ8.5）から得られるラインであるドット、両方を示しているので、この図は、分析プロセスを示しています。 初期、?...

Watch this Scientific Journal Video about Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement at JoVE.com

Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement

We describe here a relatively fast and simple approach for mapping genome-wide mammalian replication timing, from cell isolation to the basic analysis of the sequencing results. A genomic map of a representative replication program will be provided following the protocol.

DNA含有量測定法による哺乳類の複製タイミングのゲノムワイドな決意

Replication of the genome occurs during S phase of the cell cycle in a highly regulated process that ensures the fidelity of DNA duplication. Each genomic ...

The overall goal of this procedure, is to map the genome-wide replication timing program of a cell of choice. This method can help answer key questions in the replication field. Such as, how the genomic replication program changes in health and disease.The main advantage of this technique is that it is a fast and simple way to map replication timing in various cells, and conditions. The man writing the fax procedure would be Dan Lehmann, a fax expert in The Faculty of Medicine. This procedure usually starts with at least one to two times ten to the sixth fast growing unsynchronized cells.For adherent cells, aspirate and wash each plate with three milliliters of PBS without calcium of magnesium. Discard the PBS and add one milliliter of commercial Trypsin-EDTA to each plate. Incubate the cells for five minutes, at 37 degrees Celsius, until they detach.Add three milliliters of culture media to neutralize the Trypsin and collect the cells in a 15 milliliter conical tube or a five milliliter polystyrene tube. Keep the cells on ice. To begin theisprocedure, centrifuge the cells at 300 x G for five minutes at four degrees Celsius.Aspirate and wash the cells with one milliliter of cold PBS. Then, spin down the cells. After the second wash, remove the PBS and resuspend the cells in 250 microliters of cold PBS.While gently vortexing the tube, slowly add drop wise 800 microliters of minus 20 degree Celsius, 100 percent high purity ethanol. This leads to our final ethanol concentration of 70 to 80 percent. Incubate the cells on ice for 30 minutes.Begin this procedure by centrifuging the fixed cells at 500 x G for ten minutes at 4 degrees Celsius. Aspirate the supernatant carefully and wash the cells twice with one milliliter of cold PBS. Spin down after each wash.After the second wash, aspirate the supernatant and resuspend each cell with the following PI mixture. One milliliter of PBS containing five microliters of ten milligram per milliliter RNAas, and 50 microliters of one milligram per milliliter propidium iodide. Filter the sample through a 35 micrometer mesh into a 5 milliliter polystyrene tube.Seal the tube with parafilm. Incubate at room temperature for 15 to 30 minutes. Keep the samples in the dark as propidium iodide is light sensitive.The cells will be sorted using a fax machine. Use the 561 nanometer laser to differentiate cells based on their propidium iodide intensity. For optimal results, use the smallest nozzle recommended for the specific cell size.For most cells, it is 85 micrometers. Use a steady, slow flow. Usually up to 300 to 500 events per second.Using Gating, discriminate dead cells and subcellular debris by plotting FCS versus SSC. From the viable cells, discriminate doublets by plotting SSC width versus SSC height, followed by an FFC width versus FFC height plot, and a propidium iodide width versus propidium iodide height plot. For the viable signal cells, draw a histogram of the propidium iodide area intensity, which represents the DNA content of the cells.Sort cells in cold conditions into G1 and S phases. Gating for S should be wide and intrude into the G1 and G2 phases. While G1 gating should be narrow and as far from S as possible.For come cell types, it may be hard to get a nice cell cycle distribution. It is most critical to take the G1 phase without S phase cells. Remove tubes with sorted cells from the fax machine.Keep tubes on ice following the sort. Following the cell sorting, the DNA for each sample is purified using a commercial DNA purification kit following the manufacturer's directions. Next, sheer the DNA with a focused ultrasonicator to an average target peak size of 250 base pairs.Verify the size of the sheer DNA by electroforesis. The recommended size distribution is 200 to 700 base pairs with a peak at about 250 base pairs. Subsequently, the library is prepared, sequenced, and analyzed as described in the text protocol.A time of replication map for the entire chromosome one from mouse embryonic fibroblasts is shown. With a closer look at a region of approximately 50 megabases. The dots represent the normalized S to G1 ratio for individual windows, and the solid line represents the results from the cubic smoothly and interpolation.High S to G1 values corresponds to early replication, whereas low values correspond to late replication. The red arrows point to examples of constant time of replication regions, and the green arrows point to temporal transition regions. This figure shows the reproducibility of time of replication maps between triplicates of mouse embryonic fibroblasts, compared to triplicates of mouse pre-B cells in the same 18 megabase region on mouse chromosome one.The orange arrows, point to examples of differential regions between the cell lines. A heat map of the spearman correlations between the six samples further confirm the results. Once mastered, this technique from the collection of cells to the generation of maps, can be completed in one week if performed properly.After watching this video, you should have a good understanding of how to generate time of replication genomic maps for growing cells.

Research

JoVE Journal

Genetics

In Situ Labeling of Mitochondrial DNA Replication in Drosophila Adult Ovaries by EdU Staining

In Situ Labeling of Mitochondrial DNA Replication in Drosophila Adult Ovaries by EdU Staining

EDU染色によりショウジョウバエアダルト卵巣内のミトコンドリアDNA複製のその場の標識で

The mitochondrial genome is inherited exclusively through the maternal line. Understanding of how the mitochondrion and its genome are proliferated and ...

遺伝学

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins

遅行本鎖DNA合成の動態<em&gt;インビトロ</em&gt;バクテリオファージT7複製タンパク質によって

Here we provide protocols for the kinetic examination of lagging-strand DNA synthesis in vitro by the replication proteins of bacteriophage T7. The T7 ...

Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae

Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae

ChEC-seqとのタンパク質 - DNA相互作用のゲノムワイドマッピング<em&gt; Saccharomyces cerevisiae</em

Genome-wide mapping of protein-DNA interactions is critical for understanding gene regulation, chromatin remodeling, and other chromatin-resident ...

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Determination of the Optimal Chromosomal Locations for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

新規トランスポゾンを介したアプローチを使用してエシェリヒア属大腸菌DNA 要素の最適な染色体の場所の決定

The optimal chromosomal position(s) of a given DNA element was/were determined by transposon-mediated random insertion followed by fitness selection. In ...

Mapping Genome-wide Accessible Chromatin in Primary Human T Lymphocytes by ATAC-Seq

ATAC Seq による主なヒト T リンパ球のゲノム広いアクセス可能なクロマチンをマッピング

Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) is a method used for the identification of open (accessible) regions ...

Determining Genome-wide Transcript Decay Rates in Proliferating and Quiescent Human Fibroblasts

ひと線維芽細胞の増殖静止ゲノムワイドな転写減衰率を決定します。

Quiescence is a temporary, reversible state in which cells have ceased cell division, but retain the capacity to proliferate. Multiple studies, including ...

Generation of Genome-wide Chromatin Conformation Capture Libraries from Tightly Staged Early Drosophila Embryos

Generation of Genome-wide Chromatin Conformation Capture Libraries from Tightly Staged Early Drosophila Embryos

しっかりと段階的な初期のショウジョウバエの胚からゲノムワイドなクロマチン構造キャプチャ ライブラリの生成

Investigating the three-dimensional architecture of chromatin offers invaluable insight into the mechanisms of gene regulation. Here, we describe a ...

Profiling DNA Replication Timing Using Zebrafish as an In Vivo Model System

Profiling DNA Replication Timing Using Zebrafish as an In Vivo Model System

In Vivoモデル システムとしてゼブラフィッシュを用いた DNA 複製タイミングのプロファイリング

DNA replication timing is an important cellular characteristic, exhibiting significant relationships with chromatin structure, transcription, and DNA ...

Promoter Capture Hi-C: High-resolution, Genome-wide Profiling of Promoter Interactions

プロモーター キャプチャこんにちは-c: 高解像度, ゲノム プロモーターの相互作用のプロファイリング

The three-dimensional organization of the genome is linked to its function. For example, regulatory elements such as transcriptional enhancers control the ...

A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells

シンプル、迅速、かつ定量的アッセイのプラスミド dna-タンパク質クロスリンクのメジャー修理に哺乳類細胞にトランスフェクション

The purpose of this method is to provide a flexible, rapid, and quantitative technique to examine the kinetics of DNA-protein crosslink (DPC) repair in ...