Illumina測序文庫構(gòu)建
ThruPLEX® DNA-seq Kit
· 高靈敏度、高性能：以低至50 pg至50 ngDNA為起始材料
· 完整的文庫構(gòu)建試劑：優(yōu)化的檢索引物和接頭試劑、酶、緩沖液及無核酸酶水
· 快速簡便的工作流程：單管或單孔3步操作，2小時，無需轉(zhuǎn)移操作
· 兼容主流目標(biāo)區(qū)域富集平臺：Agilent SureSelect®, Roche Nimblegen® SeqCap

詳細(xì)介紹

Rubicon RB4406/R400406 ThruPLEX DNA-seq 48D Kit DNA測序試劑盒紅榮微再*代理基因測序試劑盒

美國Rubicon Genomics---單細(xì)胞測序文庫構(gòu)建的

1. 二代測序建庫試劑（微量起始樣品 50pg - 50ng）

2. 單細(xì)胞擴增和單細(xì)胞測序建庫試劑（廣泛應(yīng)用于生殖領(lǐng)域，是該領(lǐng)域的金標(biāo)準(zhǔn)。）

3. 全RNA擴增試劑（微量起始樣品，可以低至10個細(xì)胞的總RNA）

Rubicon Genomics成立于2000年，總部位于美國密歇根州。該公司的核心競爭力產(chǎn)品包括酶學(xué)、核酸化學(xué)、樣品的加工生產(chǎn)，該公司致力于創(chuàng)新、知識產(chǎn)權(quán)的發(fā)展和持續(xù)的產(chǎn)品開發(fā)。

Rubicon Genomics開發(fā)創(chuàng)新、高品質(zhì)的、核酸文庫的制備及配套產(chǎn)品，RB4406/R400406，能夠?qū)εR床樣本進行簡單、快速可靠和高靈敏度的分析。

Rubicon Genomics，總部位于美國密歇根州，產(chǎn)品主要包括以下幾類：

【主要產(chǎn)品】

1. ThruPLEX® Plasma-seq Kit

2. ThruPLEX® DNA-seq Kit

3. PicoPLEX® DNA-seq Kit

4. PicoPLEX® WGA Kit

5. TransPLEX® C-WTA Kit

【Rubicon RB4523/R400523 ThruPLEX DNA-seq 6S (12 )Kit DNA測序試劑盒產(chǎn)品應(yīng)用】

1. 外顯子組富集

2. SureSelect® 富集

3. SeqCap® EZ 富集

4. 福爾馬林固定石蠟包埋DNA測序

5. 胚胎植入前遺傳學(xué)篩查(PGS）/種植前遺傳學(xué)診斷（PGD）

6. DNA測序

7. RNA測序

8. 染色體免疫共沉淀測序

歡迎您致電華雅再生醫(yī)學(xué)旗艦公司：紅榮微再（上海）生物工程技術(shù)有限公司 ：152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348

提供產(chǎn)品：華雅再生醫(yī)學(xué)旗艦公司-紅榮微再（上海）生物工程技術(shù)有限公司為中國再生醫(yī)學(xué)和生物工程技術(shù)領(lǐng)域的組織器官再生研究、組織器官功能重建研究、干細(xì)胞技術(shù)研究、組織工程生物支架材料研究、器官移植醫(yī)療器械產(chǎn)品等再生醫(yī)學(xué)產(chǎn)業(yè)鏈的相關(guān)客戶提供：iPS/ES胚胎干細(xì)胞的種子細(xì)胞、心肌細(xì)胞、人源肝臟細(xì)胞、MSC間充質(zhì)干細(xì)胞、基底膜基質(zhì)膠細(xì)胞支架、干細(xì)胞生長分化調(diào)控因子、干細(xì)胞藥物研究的小分子抑制劑、調(diào)節(jié)劑、小分子化合物、功能膠原蛋白生物材料、3D細(xì)胞培養(yǎng)基質(zhì)膠、器官模型3D打印技術(shù)材料、神經(jīng)干細(xì)胞培養(yǎng)基、STEM121抗體、BD科研P100、P800蛋白質(zhì)組學(xué)采血管、BD血漿準(zhǔn)備管PPT管、BD PAXgene全血RNA樣本收集管、BD公司CPT/PRP美容采血管、Streck Cell-Free RNA BCT游離RNA樣本收集管、Streck Cell-Free DNA Urine Preserve游離DNA尿液保存管、Streck Cell-Free DNA BCT游離DNA采血管、循環(huán)腫瘤細(xì)胞CTC及*產(chǎn)前基因檢測采血管、illumina高品質(zhì)NGS文庫制備試劑盒、ThruPLEX ctDNA測序試劑盒、全基因組測序、全外顯子測序、靶向測序、ChIP-Seq測序、CNV和染色體異倍性分析、Rubicon Genomics單細(xì)胞全基因組測序試劑盒、生物醫(yī)學(xué)試劑耗材儀器等種類豐富的產(chǎn)品，以“Focus on Tissue and Organ Regeneration專注組織器官再生與功能重建”為己任，我們與客戶攜手為中國再生醫(yī)學(xué)產(chǎn)業(yè)的發(fā)展而前行。

【Rubicon Genomics公司基因測序試劑盒產(chǎn)品詳情】

產(chǎn)品名稱	平臺	應(yīng)用程序	樣本	樣本量	技術(shù)
ThruPLEX®DNA-seq Kit	Illumina NGS	WGS, WES, 靶向測序，染色體免疫共沉淀測序, 轉(zhuǎn)錄組測序	FFPE DNA, gDNA, cDNA, Cell-free DNA, ChIP DNA, amplicons	50 pg – 50 ng	高靈敏度基于連接的擴增
ThruPLEX®-FD Prep Kit	Illumina NGS	WGS, WES, 靶向測序，ctDNA分析	Cell-free DNA isolated from plasma	<1 ng – 30 ng	高靈敏度基于連接的擴增
PicoPLEX™ DNA-seq Kit	Illumina NGS	CNV,Aneuploidy	Eukaryotic Cells or gDNA	<6 pg – 60pg	準(zhǔn)隨機引物線性擴增
PicoPLEX™ WGA Kit	陣列，PCR	CNV,Aneuploidy	Eukaryotic Cells or gDNA	<6 pg – 60 pg DNA	準(zhǔn)隨機引物線性擴增
TransPLEX®C-WTA Kit	陣列，PCR	基因表達分析	RNA	10 – 300 ng RNA	準(zhǔn)隨機引物擴增

Rubicon RB4406/R400406 ThruPLEX DNA-seq 48D Kit DNA 基因測序試劑盒的綜合介紹

Illumina測序文庫構(gòu)建

ThruPLEX^® DNA-seq Kit

· 高靈敏度、高性能：以低至50 pg至50 ngDNA為起始材料
· 完整的文庫構(gòu)建試劑：優(yōu)化的檢索引物和接頭試劑、酶、緩沖液及無核酸酶水
· 快速簡便的工作流程：單管或單孔3步操作，2小時，無需轉(zhuǎn)移操作
· 兼容主流目標(biāo)區(qū)域富集平臺：Agilent SureSelect^®, Roche Nimblegen^® SeqCap^® EZ 及 IDT xGen^® Lockdown^® probes

■ 產(chǎn)品說明：

ThruPLEX DNA-seq Kit 基于新的ThruPLEX 技術(shù)，可以低至50 pg的DNA起始構(gòu)建DNA文庫，在提高靈敏度的同時可以構(gòu)建多達96種檢索引物文庫。本試劑適用于多種樣品來源的片段化DNA樣品，基因組DNA或體液樣品如游離DNA，F(xiàn)FPE樣品來源DNA和cDNA。ThruPLEX DNA-seq Kit 整體流程可以在單管或單孔中進行，約2個小時即可完成，無需純化或樣品轉(zhuǎn)移步驟。試劑盒提供12、48或96種Illumina單端或雙端檢索引物，該系列引物保存在反應(yīng)管中(6或12)或預(yù)分裝密封于條形碼標(biāo)記的微孔板中。ThruPLEX DNA-seq Kit 可以用于DNA-seq，RNA-seq，ChIP-seq，并且可以兼容主流平臺實現(xiàn)強有力的目標(biāo)區(qū)域富集。

圖1. ThruPLEX DNA-seq Kit單管操作流程。

圖2. ThruPLEX DNA-seq Kit技術(shù)概要。ThruPLEX技術(shù)以片段化雙鏈DNA起始(0.05 ng至50 ng)只需3步反應(yīng)，簡單高效。

圖3. ThruPLEX DNA-seq Kit 優(yōu)于其他NGS文庫制備試劑。與其他試劑相比，ThruPLEX DNA-seq Kit可以更少的起始DNA，構(gòu)建多樣性更好（左圖），重復(fù)更少（右圖）的文庫（25 M read pairs）。NGS文庫分別采用圖示對照gDNA起始量，以相應(yīng)的文庫制備試劑制備，采用MiSeq v3 flow cell測序。
*數(shù)據(jù)來源于Takara Bio Inc.

圖4. ThruPLEX DNA-seq Kit 減少了GC偏差。以人類基因組相對覆蓋度為GC含量（%）的函數(shù)（50 ng DNA起始）。（左圖）ThruPLEX DNA-seq Kit 在整個范圍內(nèi)都有出色的覆蓋。（右圖）ThruPLEX DNA-seq Kit （金色）與參試的其他試劑一樣好或更好。請注意，在這些圖中的覆蓋度應(yīng)該為1。人類基因組中的GC含量分布以紅色柱形圖表示。
*數(shù)據(jù)來源于Takara Bio Inc.

圖5. ThruPLEX DNA-seq Kit的起始量更低、范圍更廣。與其他試劑相比，ThruPLEX DNA-seq Kit的起始量靈敏度是其他試劑的20倍，起始量范圍從50 pg至50 ng，無需流程或接頭濃度優(yōu)化，并且包含檢索引物和接頭。

圖6. ThruPLEX DNA-seq Kit工作流程優(yōu)勢：2小時即可獲得高質(zhì)量NGS文庫。ThruPLEX DNA-seq Kit將雙鏈DNA制備成檢索引物文庫只需3步：末端修復(fù)，接頭連接，及高保真文庫擴增。簡化的工作流程，在單管或單孔中2個小時即可完成，避免樣品損失及操作誤差。

■ ThruPLEX DNA-seq Kit產(chǎn)品組成

· Template Preparation Buffer（Red）
· Template Preparation Enzyme（Red）
· Library Synthesis Buffer（Yellow）
· Library Synthesis Enzyme（Yellow）
· Library Amplification Buffer（Green）
· Library Amplification Enzyme（Green）
· Nuclease-Free Water（Clear）
· Indexing Reagent（Blue）or Indexing Plate

■ ThruPLEX DNA-seq Kit保存 －20℃

2017年1月，Rubicon Genomics，Inc.成為Takara Bio的一員，現(xiàn)在寶日醫(yī)生物技術(shù)（北京）有限公司（Takara Bio北京分公司）開始在國內(nèi)銷售Rubicon公司開發(fā)的下一代測序試劑。Rubicon公司成立于2000年，公司總部位于美國密歇根州。Rubicon公司開發(fā)的產(chǎn)品主要基于“ThruPLEX®技術(shù)”和“PicoPLEX®技術(shù)”，可以微量DNA起始構(gòu)建下一代測序文庫。ThruPLEX®技術(shù)”可從血漿中的微量游離DNA構(gòu)建高多樣性文庫，準(zhǔn)確檢測低豐度DNA變異，用途廣泛。“PicoPLEX®技術(shù)”，可從單細(xì)胞等微量DNA起始構(gòu)建高重復(fù)再現(xiàn)性文庫，用于拷貝數(shù)變異分析和染色體異倍性分析。Rubicon產(chǎn)品豐富了Takara基于SMART®技術(shù)的下一代測序產(chǎn)品線，為科研工作者在基因組學(xué)、單細(xì)胞和臨床應(yīng)用研究等領(lǐng)域提供更多解決方案。代表產(chǎn)品之一為：Rubicon Genomics RB3050/R30050 PicoPLEX® WGA Kit 單細(xì)胞全基因組擴增試劑盒Whole Genome Amplification from Single Cells。

2016年年底，TAKARA寶日醫(yī)生物技術(shù)（北京）有限公司的市場部部長與寶生物工程（大連）有限公司的銷售部部長分別從北京和大連飛往上海，對華雅再生醫(yī)學(xué)的旗艦營銷公司：紅榮微再（上海）生物工程技術(shù)有限公司進行了實地考察，雙方深入地探討了未來干細(xì)胞及再生醫(yī)學(xué)市場發(fā)展的趨勢和方向，在雙方公司中長期發(fā)展戰(zhàn)略上，志同道合地看好再生醫(yī)學(xué)市場的發(fā)展前景，歷史性的醫(yī)療革命是我們這代人肩負(fù)的偉大使命！

TAKARA公司日本總部是：“iPS干細(xì)胞之父”諾貝爾獎獲得者的故鄉(xiāng)，具有zui前沿的iPS干細(xì)胞技術(shù)實力，TAKARA公司旗下的Cellartis子公司，擁有超過40人的研發(fā)團隊，擁有超過15年的人干細(xì)胞研究經(jīng)驗，建立并儲存大量的人iPS和ES細(xì)胞系。TAKARA旗下的Stem Cells公司SC Proven系列：憑借20多年干細(xì)胞技術(shù)經(jīng)驗研發(fā)，包括干細(xì)胞研究用培養(yǎng)基、抗體等耗材，具有龐大的客戶群(美國、歐洲、日本)。

2017年TAKARA寶生物日本總部又收購了：美國Rubicon Genomics單細(xì)胞測序文庫構(gòu)建的，代表性產(chǎn)品之一為：Rubicon Genomics RB3050/R30050 PicoPLEX® WGA Kit 單細(xì)胞全基因組擴增試劑盒Whole Genome Amplification from Single Cells，ThruPLEX® Plasma-seq Kit，ThruPLEX® DNA-seq Kit，PicoPLEX® DNA-seq Kit，PicoPLEX® WGA Kit， TransPLEX® C-WTA Kit.

ThruPLEX DNA-seq kit可以使用什么樣的樣品和投入？

ThruPLEX DNA-seq試劑盒設(shè)計用于各種各樣的DNA樣品，其量在50pg和50ng之間變化。為了確保實驗提供高品質(zhì)的數(shù)據(jù)，下表提供了一系列的依據(jù)（1）應(yīng)用程序和（2）的源材料的人DNA的推薦的輸入量。您選擇使用的實際數(shù)量可能與以下建議的不同。注意，隨著輸入DNA的量減少，文庫的多樣性也可能降低

應(yīng)用輸入樣品量（50 - 500 bp ds DNA）推薦輸入量

WGS，WES，SNV，indels，SNP，STR gDNA 10 - 50 ng

FFPE DNA 100 ng

血漿DNA 1 - 50 ng

CNV血漿，gDNA或FFPE DNA 50 pg - 100 ng *

ChIP-seq ChIP DNA 50 pg - 50 ng *

RNA-seq cDNA 50 pg - 50 ng *

*科學(xué)出版物中引用少于50 pg的用途。

ThruPLEX DNA-seq Kit是否與任何目標(biāo)濃縮系統(tǒng)兼容？

是的，ThruPLEX DNA-seq的套件是與主要外顯子組和靶向富集產(chǎn)品，包括安捷倫SureSelect®，羅氏NimbleGen®SeqCap®EZ和自定義面板兼容。 ThruPLEX DNA-seq Kit目標(biāo)富集方案和應(yīng)用筆記可以在這里訪問。

我購買了一種ThruPLEX DNA-seq 48D試劑盒，一次使用4-5個樣品。為什么我用完試劑？

ThruPLEX DNA-SEQ試劑盒的試劑包含足夠的體積，以制備主混合物四次，隨著時代的索引試劑應(yīng)使用的號碼*（zui多4個冷凍/解凍循環(huán)）。當(dāng)訪問套件超過四次時，我們不能保證有足夠的試劑量可用。

ThruPLEX DNA-seq是否有足夠的過量填充以在我的自動化設(shè)備上使用？

每個自動化平臺都有自己的死體積和試劑要求;因此，ThruPLEX DNAs-seq Kit中試劑的體積可能不能提供手動進行的預(yù)期反應(yīng)數(shù)。

剪切后，我需要對DNA樣本進行清理嗎？

沒有，只是繼續(xù)到ThruPLEX DNA-seq的試劑盒使用手冊中的模板制備步驟（第直噴），以維護您的總DNA和圖書館的整體多樣性。

ThruPLEX DNA-seq Kit可以使用什么樣的DNA樣本？

分離的雙鏈DNA，如：

機械剪切DNA

酶切片段DNA

來自血漿，尿液和其他生物流體的低分子量無細(xì)胞DNA

雙鏈cDNA

染色質(zhì)免疫沉淀（ChIP）DNA

來自福爾馬林固定材料（FFPE）的超聲處理DNA

RNA可以與ThruPLEX DNA-seq Kit一起使用嗎？

是的，但是在使用ThruPLEX DNA-seq Kit制備之前，需要將RNA轉(zhuǎn)化為雙鏈cDNA并將其片段化至適當(dāng)?shù)拇笮　?/span>

血漿，尿液或其他生物流體可以直接與ThruPLEX DNA-seq Kit一起使用嗎？

不，模板DNA必須首先用DNA提取/純化試劑盒提取。由于來自這些來源的DNA通常被降解/分裂，所以不需要額外的分裂。樣品中使用的體積

ThruPLEX DNA-seq Kit必須不超過10μL。

我可以為單端和雙端NGS測序準(zhǔn)備樣品嗎？

是的，ThruPLEX DNA-seq Kit與單端和雙向序列測序兼容。zui終擴增的文庫將包含與Illumina相容的索引序列。

ThruPLEX DNA-seq Kit是否需要DNA樣本輸入體積？

是的，所需的DNA樣品量為10μL。

出版物：

無細(xì)胞DNA包含通知其原始組織的體內(nèi)核小體足跡。 Snyder等。

Overview

ThruPLEX® DNA-seq Kit builds on the innovative ThruPLEX chemistry to generate DNA libraries from as little as 50 pg of DNA and increases sensitivity while providing up to 96 indexes for multiplexing. This kit can be used with fragmented DNA from any sample source – genomic, biofluids such as cell-free DNA, DNA from FFPE materials and cDNA. The entire ThruPLEX DNA-seq Kit workflow is performed in a single tube or well in about 2 hours and requires no purification steps or sample transfers. Kits contain 12, 48 or 96 single- or dual-read Illumina-compatible indexes either in tubes (6 or 12) or pre-dispensed and sealed in barcoded microplates. ThruPLEX DNA-seq Kit can be used in any DNA-seq application, RNA-seq, ChIP-seq and offers robust target enrichment performance with all of the leading platforms.

ThruPLEX DNA-seq Kit is protected by U.S. Patents 7,803,550; 8,071,312; 8,399,199; 8,728,737 and corresponding foreign patents. Additional patents are pending. For Research Use only.

Rubicon RB4406/R400406 ThruPLEX DNA-seq 48D Kit DNA 基因測序試劑盒的優(yōu)點優(yōu)勢

ADVANTAGES

High performance from lowest input: 50 pg to 50 ng

Complete package for creating libraries: optimized indexing and adaptor reagents, enzymes, buffers, and nuclease-free water

Fast and simple workflow: 3 steps in a single tube or well in about 2 hours, no transfers necessary

Compatible with major target enrichment platforms: Agilent SureSelect®, Roche Nimblegen® SeqCap® EZ, and IDT xGen® Lockdown® probes

Rubicon ThruPLEX DNA-seq 48D Kit DNA 基因測序試劑盒產(chǎn)品規(guī)格

SPECIFICATIONS

Store at -20 °C.

Guaranteed for 9 months at -20 °C in a constant temperature freezer.

Ordering Information

CAT. NO. R400406 ThruPLEX DNA-seq 48D Kit (48 reactions, 48 dual indexes)

CAT. NO. R400407 ThruPLEX DNA-seq 96D Kit (96 reactions, 96 dual indexes)

CAT. NO. R400427 ThruPLEX DNA-seq 48S Kit (48 reactions, 48 single indexes)

CAT. NO. R400428 ThruPLEX DNA-seq 12S (48) Kit (48 reactions, 12 single indexes)

CAT. NO. R400523 ThruPLEX DNA-seq 6S (12) Kit (12 reactions, 6 single indexes)

ThruPLEX DNA-seq contains everything needed, including indexes in a barcoded 96-well plate.

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Rubicon ThruPLEX DNA-seq 48D Kit DNA 基因測序試劑盒產(chǎn)品信息

RESOURCES

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ThruPLEX DNA-seq Kit Instruction Manual

ThruPLEX DNA-seq Product Sheet

ThruPLEX DNA-seq Dual Index Quick Protocol

ThruPLEX DNA-seq Dual Index Plate Guide

ThruPLEX DNA-seq Single Index Quick Protocol

ThruPLEX DNA-seq Single Index Plate Guide

ThruPLEX DNA-seq Index Sequences 6S (CSV)

ThruPLEX DNA-seq Index Sequences 12S (CSV)

ThruPLEX DNA-seq Index Sequences 48S (CSV)

ThruPLEX DNA-seq SDS

ThruPLEX DNA-seq Kit Outperforms Other NGS LIbrary Prep Kits

ThruPLEX DNA-seq Kit yielded more diverse libraries (left) with less input DNA and generated fewer duplicates (right) at 25M read pairs than other kits. NGS libraries were prepared with the indicated amount of reference gDNA using the library preparation kits indicated and sequenced on a MiSeq, v3 flow cell.

ThruPLEX DNA-seq Kit Single-Tube Workflow

Starting with as little as 50 pg of fragmented double-stranded DNA or cDNA, ThruPLEX DNA-seq Kit creates indexed libraries in 3 simple steps: end repair, adapter ligation, and high-fidelity library amplification. The streamlined workflow prevents sample loss and is performed in about 2 hours in a single tube or well preventing sample loss and enhancing positive sample identification.

ThruPLEX DNA-seq Kit Minimizes GC Bias

Relative coverage of the human genome as a function of %GC (50 ng input DNA). (Left) ThruPLEX DNA-seq Kit has excellent coverage throughout the entire range. (Right) ThruPLEX DNA-seq Kit (gold) performs as well as or better than other kits tested. Note that the optimal coverage should be equal to 1 in these graphs. GC composition distribution of the human genome is represented by the red bars.

ThruPLEX DNA-seq Kit Has a Lower and Broader Input Range

ThruPLEX DNA-seq Kit has a lower and broader recommended input range than other kits. The sample input amount for ThruPLEX DNA-seq Kit is 20 times more sensitive than other kits, ranging from 50 pg and 50 ng and requires no optimization of the protocol or adapter concentration. Indexes and adapters are included with the kit.

ThruPLEX DNA-seq Kit Workflow Advantage: High Quality NGS Libraries in About 2 Hours

ThruPLEX DNA-seq Kit converts double-stranded DNA samples to indexed libraries in 3 simple steps: end repair, adapter ligation, and high-fidelity library amplification. The streamlined workflow prevents sample loss and handling errors and is performed in a single tube or well in less than 2 hours.

ThruPLEX DNA-seq Kit Technology

ThruPLEX technology is a 3-step reaction that starts with fragmented double-stranded DNA (0.05 ng to 50 ng) which is repaired in a highly efficient process. Background is reduced using double-stranded adaptors with no single-stranded tails. Blunt end ligation occurs with high-efficiency. Blocked 5’ends reduce adaptor-adaptor ligation. Background is further reduced by destroying unused adaptors after ligation

FAQS

Click here to read all FAQ’s on ThruPLEX DNA-seq Kit

1. What types of samples and inputs can be used with the ThruPLEX DNA-seq kit?

ThruPLEX DNA-seq Kit is designed to work on a wide variety of DNA samples with amounts varying between 50 pg and 50 ng. To ensure that your experiment delivers quality data, the table below provides a range of recommended input amounts of human DNA based on (1) your application and (2) the source material. The actual amount that you chose to use may vary from those suggested below. Note that as the amount of input DNA decreases, the diversity of the library may also decrease

Application Input sample amount (50 – 500 bp ds DNA) Recommended input amount

WGS, WES, SNV, indels, SNP, STR gDNA 10 – 50 ng

FFPE DNA 100 ng

plasma DNA 1 – 50 ng

CNV plasma, gDNA or FFPE DNA 50 pg – 100 ng*

ChIP-seq ChIP DNA 50 pg – 50 ng*

RNA-seq cDNA 50 pg – 50 ng*

*Use of less than 50 pg has been cited in scientific publications.

2. Is ThruPLEX DNA-seq Kit compatible with any target enrichment systems?

Yes, ThruPLEX DNA-seq Kit is compatible with the major exome and target enrichment products, including Agilent SureSelect®, Roche NimbleGen® SeqCap® EZ and custom panels. ThruPLEX DNA-seq Kit target enrichment protocols and application notes can be accessed here.

3. I purchased a ThruPLEX DNA-seq 48D kit and have been using it for 4-5 samples at a times. Why am I running out of reagents?

ThruPLEX DNA-seq Kit reagents contain enough volume to prepare master mixes FOUR times, coincident with the number of times the indexing reagents should be used (up to 4 freeze/thaw cycles). When accessing the kit more than the four times, we cannot guarantee that enough reagent volume will be available.

4. Does ThruPLEX DNA-seq have enough overfill to be used on my automation device?

Each automation platform has its own dead volume and reagent requirements; hence the volume of reagents in ThruPLEX DNAs-seq Kit may not provide the expected number of reactions that would be performed manually.

5. Do I need to perform cleanup on my DNA sample(s) after shearing?

No, simply proceed to the Template Preparation Step (Section D.I.) in the ThruPLEX DNA-seq Kit Instruction Manual in order to preserve your total DNA and the overall diversity of the library.

6. What types of DNA samples can be used with ThruPLEX DNA-seq Kit?

Fragmented, double-stranded DNA, such as that from:

Mechanically-sheared DNA

Enzymatically-fragmented DNA

Low molecular weight cell-free DNA from plasma, urine, and other biofluids

Double-stranded cDNA

Chromatin Immunoprecipitated (ChIP) DNA

Sonicated DNA from formalin-fixed material (FFPE)

7. Can RNA be used with ThruPLEX DNA-seq Kit?

Yes, but RNA needs to be converted into double-stranded cDNA and fragmented to proper size prior to preparation with ThruPLEX DNA-seq Kit.

8. Can plasma, urine, or other biofluids be directly used with ThruPLEX DNA-seq Kit?

No, the template DNA must be first extracted with a DNA extraction/purification kit. Since DNA from these sources is normally degraded/fragmented, additional fragmentation is unnecessary. The volume of the sample used in the

ThruPLEX DNA-seq Kit must be no more than 10 µL.

9. Can I prepare samples for both single- and paired-end NGS sequencing?

Yes, ThruPLEX DNA-seq Kit is compatible with both single- and paired-end sequencing. The final amplified libraries will contain Illumina-compatible index sequences.

10. Is there a required DNA sample input volume for ThruPLEX DNA-seq Kit?

Yes, the required DNA sample volume is 10 µL.

Publication:

Cell-free DNA Comprises an In Vivo Nucleosome Footprint that Informs Its Tissues-of-Origin. Snyder et all.

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