當(dāng)前位置 ： NEB >>> NEB/NEBNext FFPE DNA Repair v2 Module/24 reactions/E7360S
NEB/NEBNext FFPE DNA Repair v2 Module/24 reactions/E7360S
  • NEB/NEBNext FFPE DNA Repair v2 Module/24 reactions/E7360S

NEB/NEBNext FFPE DNA Repair v2 Module/24 reactions/E7360S

價(jià)格: 試用 市場(chǎng)價(jià): 0.00
貨號(hào): E7360S
品牌: NEB
規(guī)格
數(shù)量
庫(kù)存(0)
特別 提示
代購(gòu)產(chǎn)品：無(wú)質(zhì)量問(wèn)題不接受退換貨，下單前請(qǐng)仔細(xì)核對(duì)信息。
下單后請(qǐng)及時(shí)聯(lián)系客服核對(duì)商品價(jià)格，訂單生效后再付款。
資深產(chǎn)品顧問(wèn)
咨詢顧問(wèn)

全國(guó)免費(fèi)服務(wù)熱線

4000-520-616

  • 自營(yíng)商城 一站式服務(wù)
  • 廠家直采 剔除溢價(jià)
  • 品質(zhì)甄選 正品保證
  • 嚴(yán)控流程 只做188精品
  • 極速物流 如約送貨
  • 詳情
  • 使用說(shuō)明
  • 常見(jiàn)問(wèn)題

    • The NEBNext FFPE DNA Repair v2 Module contains enzymes and buffers that have been optimized to repair FFPE DNA in next generation sequencing workflows in a streamlined manner.The NEBNext FFPE DNA Repair v2 Module improves upon the performance of the original NEBNext FFPE DNA Repair Mix:

      • Higher repair efficiency with FFPE DNA
      • A more streamlined workflow for NGS library preparation
      • A more convenient reaction buffer containing all required buffer components for both efficient FFPE DNA repair and downstream end repair and dA-tailing.
      • No cleanup is required between repair and library prep, through the use of Thermolabile Proteinase K.

      Table 1: Types of FFPE DNA damage and their ability to be repaired by the NEBNext FFPE DNA Repair v2 Module

      FFPE DAMAGE TYPE REPAIRED BY THE FFPE DNA REPAIR v2 MODULE
      Deamination of cytosine to uracil Yes
      Nicks and gaps Yes
      Oxidized bases Yes
      Blocked 3´ ends Yes
      DNA fragmentation No
      DNA-protein crosslinks No
      Figure 1: Workflow demonstrating the use of the NEBNextFFPE DNA Repair v2 Module with the NEBNext Ultra™ II DNA Library Prep Kit for Illumina®
      Figure 2: The NEBNext FFPE DNA Repair v2 Module enables robust library preparation from a broad range of FFPE DNA sample qualities.
      Libraries were prepared with 25 ng of Covaris®acoustic-sheared FFPE DNA samples of different qualities and tissue sources. The NEBNext FFPE DNA Repair v2 Module was used, followed by NEBNext Ultra II DNA library preparation (NEB #E7645) with 9 PCR cycles. Libraries were quantified using the Agilent® HS D1000 TapeStation®. The NEBNext FFPE DNA Repair v2 Module improves the yield of FFPE libraries by varying degrees depending on the quality and damage types present in the input DNA. Error bars indicate the standard deviation of two replicates for each library sample.
      Figure 3: The NEBNext FFPE DNA Repair v2 Module enables robust library preparation from 5 to 250 ng of FFPE DNA input using both NEBNext hairpin and Unique Dual Index UMI Adaptors
      Libraries were prepared with 5, 50, or 250 ng of three different quality normal liver FFPE DNA samples comparing treatment with and without the NEBNext FFPE DNA Repair v2 Module. The NEBNext Ultra II DNA Library Preparation Kit (NEB #E7645) was used with (a) the NEBNext hairpin adaptor and 11, 8 and 6 PCR amplification cycles for 5, 50 and 250 ng inputs, respectively, and (b) the NEBNext Unique Dual Index UMI DNA Adaptors (NEB #E7395) and 11, 8 and 6 PCR cycles for 5, 50 and 250 ng inputs, respectively. All libraries were quantified using the Agilent HS D1000 Tapestation and the average yield of 2 libraries (5 and 50 ng) and 1 replicate (250 ng) was plotted. Error bars indicate standard deviation for the 5 and 50 ng library replicates. The NEBNext FFPE DNA Repair v2 Module is compatible with 5 to 250 ng inputs of FFPE DNA and different NEBNext adaptors.
      Figure 4: The NEBNext FFPE DNA Repair v2 Module improves library quality metrics including mapping rate, properly paired reads, and chimeric reads
      Libraries were prepared with 50 ng of three different quality normal liver FFPE DNA samples in triplicate, either untreated or treated with the NEBNext FFPE DNA Repair v2 Module before library preparation using the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645). Libraries were sequenced on the Illumina NextSeq®500. Paired-end reads were downsampled to 1 million reads and mapped to the GRCh38 human reference (RefSeq 884148) using Bowtie2 (v2.3.2). Mapped reads were analyzed with MarkDuplicates (v1.56.0) and Picard SAM/BAM alignment summary metrics (v1.56.0). Treatment with the NEBNext FFPE DNA Repair v2 Module increases the mapping rate and decreases the level of non-properly paired and chimeric reads.
      Figure 5: The NEBNext FFPE DNA Repair v2 Module repairs abundant cytosine deamination artifacts present in FFPE DNA samples
      50 ng samples of two different normal liver FFPE DNAs (DIN 2.0 and DIN 1.8) were either untreated or treated in duplicate with the NEBNext FFPE DNA Repair v2 Module, followed by library preparation using the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645). Libraries were sequenced on the Illumina NextSeq®500. Paired-end reads were downsampled to 1 million reads and mapped to the GRCh38 human reference (RefSeq 884148) using Bowtie2 (v2.3.2). Mapped reads were analyzed with MarkDuplicates (v1.56.0) and dCdT (CT) mutations were analyzed using Tasmanian (V0.1.3) according to the equation MAX(([C T] - [G A]) / ([NC]+[NG]), EXP(-10)). (a) The frequency of artifactual CT mutations was plotted as a function of read position (0 to 75 bp) in read 1 and read 2. The abundance and positional bias of these artifacts were reduced in two different FFPE DNA samples when treated with the NEBNext FFPE DNA Repair v2 Module. (b) Artifactual mutations identified in read 2 were quantified as an overall frequency. Two replicates for each condition are shown.
      Figure 6: The NEBNext FFPE DNA Repair v2 Module repairs oxidative damage in FFPE and non-FFPE DNA samples
      A. Libraries were prepared in duplicate with 25 ng of two different normal FFPE DNA samples of different qualities and tissue sources, comparing untreated to treated with the NEBNext FFPE DNA Repair v2 Module. Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645), sequenced on the Illumina NextSeq® 500, downsampled to 2 million reads, mapped to the GRCh38 human reference genome using Bowtie2 (v2.3.2). Mapped reads were analyzed by MarkDuplicates (v1.56.0) and Tasmanian (V0.1.3). The frequency of artifactual dGdT (GT) mutations in read 1 are shown for each library replicate. dGdT mutations were calculated according to the equation MAX(([G T] - [C A]) / ([NG]+[NC]), EXP(-10)).B. Libraries were prepared in duplicate using 100 ng of human genomic DNA Covaris-sheared in either water* or Tris-EDTA pH 8.0, the NEBNext FFPE DNA Repair v2 Module, and the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645). Libraries were sequenced on the Illumina NextSeq 500, downsampled to 2 million reads and analyzed for GT mutation frequency as above. *Note: Shearing DNA in water is not recommended by the manufacturer (Covaris) but was used to generate a substrate containing oxidative damage.
      Figure 7: The NEBNext FFPE DNA Repair v2 Module improves library yield compared to treatment with UDG enzyme
      Libraries were prepared in duplicate using 50 ng FFPE DNA input from 4 different normal FFPE DNAs of different tissue sources and qualities (FFPE #1-3 with DIN 2.0 and FFPE #4 with DIN 6.7). Libraries were either untreated, treated with the NEBNext FFPE DNA Repair v2 Module, or treated with UDG enzyme (NEB #M0280) before preparing libraries with the NEBNext Ultra II DNA Library Preparation Kit for Illumina (NEB #E7645). While UDG enzyme can eliminate dU bases from input DNA, the NEBNext FFPE DNA Repair v2 Module can fully repair these damage sites and improve the final library yield (Figures 2, 3, 7) and metrics (Figure 4).
      Figure 8: The NEBNext FFPE DNA Repair v2 Module reduces false positives in somatic variant calling arising from cytosine deamination artifacts
      Libraries were prepared from 100 ng of four different FFPE DNA samples of varying qualities and tissue sources in duplicate using the NEBNext FFPE DNA Repair v2 Module or NEBNext FFPE DNA Repair Mix (NEB #M6630), the NEBNext Unique Dual Index UMI Adaptors (NEB #E7395), the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645) and 10 PCR cycles. Libraries were captured using a custom cancer hotspot panel (Twist Bioscience®) and sequenced on the Illumina NextSeq 2000. All fastq files were downsampled to the same number of reads (2 X 18 million reads fresh frozen and 2 X 13 million for FFPE). Paired reads were trimmed with fastp (version 0.20.0) and mapped with BWA mem (0.7.17). Markduplicate was processed in picard (2.20.6) with UMI information. UMI information was processed in fgbio (0.8.1) to obtain consensus sequence reads. The final bam files with UMI-based consensus reads were used for somatic variant calling with strelka2 (2.9.10). Somatic variant calls are plotted according to substitution type for each condition. NEBNext FFPE DNA Repair v2 improves the efficiency of cytosine deamination repair (CT/GA) and effectively repairs oxidative damage (GT/CA).
      This product is related to the following categories:
      Next Generation Sequencing Library Preparation
    售后保障
    螞蟻淘生物188，秉承螞蟻淘一貫的嚴(yán)謹(jǐn)態(tài)度，由螞蟻淘公司專業(yè)人員負(fù)責(zé)品控、采購(gòu)、物流、銷售、售后，保障正品優(yōu)質(zhì)。以“快速好省，為科研提供好產(chǎn)品、好價(jià)格”為理念，直接鏈接原廠家，從全國(guó)各地原制造商嚴(yán)格挑選188款科研精品，剔除品牌溢價(jià)，188生物新電商，把好的產(chǎn)品帶給科研！? 力求給你最優(yōu)質(zhì)的商品。
  • Q：生物188產(chǎn)品正品保障嗎？
    A：生物188質(zhì)量把控人員具有十年的從業(yè)經(jīng)驗(yàn)，在業(yè)界享有良好的口碑；自營(yíng)商城，直接從廠家采購(gòu)， 自己的團(tuán)隊(duì)負(fù)責(zé)國(guó)際物流和清關(guān)，中間沒(méi)有第三方，所有流程嚴(yán)格把控，100%保證正品，假一罰十。

    Q：下單后可以修改訂單嗎？
    A：下單后的商品付款之前可以修改；訂單付款成功，需要聯(lián)系我們客服進(jìn)行修改；客服電話：4000-520-616

    Q：可以開(kāi)發(fā)票嗎？
    A：本網(wǎng)站所售商品都是正規(guī)清關(guān)，均開(kāi)具16%正規(guī)發(fā)票，發(fā)票金額含配送費(fèi)金額，另有說(shuō)明的除外。

    Q：商品幾天可以發(fā)貨？
    A：生物188商品，全部現(xiàn)貨銷售，付款后即可發(fā)貨,一般一周內(nèi)送達(dá)！

    Q：如何聯(lián)系商家？
    A：有任何問(wèn)題夠可以電話咨詢我們，全國(guó)24小時(shí)免費(fèi)服務(wù)熱線：4000-520-616 或聯(lián)系我們的在線客服QQ：1570468124

    Q：收到的商品少了/發(fā)錯(cuò)了怎么辦？
    A：同個(gè)訂單購(gòu)買多個(gè)商品可能會(huì)分為一個(gè)以上包裹發(fā)出，可能不會(huì)同時(shí)送達(dá)，建議查看訂單詳情是否是 部分發(fā)貨狀態(tài)；如未收到，可聯(lián)系在線客服或者致電4000-520-616。

    Q：退換貨/維修需要多長(zhǎng)時(shí)間？
    A：一般情況下，退貨處理周期為客戶收到產(chǎn)品一個(gè)月內(nèi)（以快遞公司顯示簽收時(shí)間為準(zhǔn)），包裝規(guī)格、 數(shù)量、品種不符，外觀毀損、短缺或缺陷，請(qǐng)?jiān)谑盏截?4小時(shí)內(nèi)申請(qǐng)退換貨；特殊商品以合同條款為準(zhǔn)。

何為188

極簡(jiǎn)而嚴(yán)謹(jǐn)，我們僅銷售188款生物醫(yī)學(xué)科研用品，款款都是爆款；因?yàn)樯偎跃劢?，聚焦甄選每一款產(chǎn)品，聚焦服務(wù)每一位客戶!

關(guān)注我們 :

點(diǎn)擊QQ聯(lián)系我們
生物188微信

關(guān)注188微信公眾號(hào)

獲取最新優(yōu)惠活動(dòng)通知