•使用上面概述的修改协议,可以使用Zymobiomics™DNA/RNA小型RNA小型RNA小型RNA小型RNA稳定器收集的人类粪便样品可以成功处理DNA稳定剂。•修改的协议可产生高产量和良好的DNA质量。•实时PCR证明了标准协议的修改不会影响下游分析。分离的DNA不含任何抑制剂。
1。将Zymo(DNA/RNA)盾牌(含3 mL DNA/RNA盾牌)的痰液陷阱和管道移入护士或呼吸治疗师,然后再进入吸入房间(或进行bal手术)。要求将不超过3 mL的痰液或BAL液收集到痰液中。2。在将气管吸气(或BAL)收集到痰液中后,估计收集到痰液中收集的痰液或BAL样品的体积,并且在患者的负压室中为气管吸气(或BAL)增添了相等数量或与3 mL Zymo屏蔽层混合的BAL流体(总共6毫升)。紧紧关闭容器。3。将气管抽吸物或BAL流体与患者的负压室中的7-10X混合在一起。这将使病毒失活,并且可以根据该站点的Covid Bioscecimen加工方案将样品用血液和尿液标本运输。
放大。PCR扩增被完全抑制。用于每个PCR的DNA,然后在2.0%(w/v)琼脂糖/tae/etbr凝胶中分析等效量的反应。梯子是100 bp的DNA标记(Zymo Research)。使用Zymotaq Premix(Zymo Research)进行热门PCR。使用OnESTEP™试剂盒或竞争对手去除PCR抑制剂后DNA恢复的比较。1KB梯子(Zymo Research)被稀释至各种浓度,并用QCR抑制剂去除套件或MN公司或Company MN的套件进行处理。DNA浓度,以计算每个试剂盒的恢复%。
Omega Bio Tek E.Z.N.A. 组织DNA KIT D3396-01 OMEGA BIO TEK MAG-BIND®血液和组织DNA HDQ 96 KIT M6399-00 OMAMGA BIO TEK E.Z.N.A. Kit CMG-196 Promega Wizard Genomic DNA Purification Kit A1120 Promega Wizard SV Genomic DNA Purification System A2360 Promega MagaZorb DNA Mini-Prep Kit MB1004 Promega ReliaPrep gDNA Tissue Miniprep System A2051 Promega Wizard HMW DNA Extraction Kit A2920 Promega ReliaPrep™ Blood gDNA Miniprep System A5081 Qiagen DNEasy Blood and Tissue Kit 69504 Qiagen QIAamp Fast DNA Tissue Kit 51404 Qiagen Puregene Tissue Kit 158063 Qiagen QIAGEN Genomic-tips 20/G 10223 Qiagen MagAttract HMW DNA Kit 67563 Qiagen QIAamp DNA Blood Mini Kit 51104 Qiagen Blood & Cell Culture DNA Mini Kit (25) 13323 Sigma-Aldrich Extract-N-AMP Tissue PCR Kit XNAT2-1KT Sigma-Aldrich GenElute Blood Genomic DNA Kit NA2010-1KT Takara NucleoSpin Tissue 740952.5 Takara NucleoMag Tissue 744300.1 Takara NucleoSpin® 96 DNA RapidLyse 740110.1 Takara NucleoBond HMW DNA 740160.2 Takara NucleoMag DNA Swab 744601.1 Takara NucleoSpin® Blood 740951.5 Takara NucleoMag® Blood 200 µL 744501.1 Thermo DNA Extract All Reagents Kit 4403319 Thermo MagMax DNA Multi-Sample Kit 4413020 Thermo JetFlex Genomic DNA Purification Kit A30700 Thermo GeneJet Genomic DNA Purification Kit K0721 Thermo GeneJet Genomic DNA Purification Kit K0721 Thermo PureLink Genomic DNA Mini Kit K182001 Thermo PureLink Genomic DNA Mini Kit K182001 Thermo ChargeSwitch gDNA Mini Tissue Kit CS11204 Zymo Quick-DNA MiniPrep D3024 Zymo Quick-DNA Miniprep Plus D4068 Zymo Quick-DNA magbead Plus Kit D4081 Zymo Quick-DNA HMW HMW Magbead d6060Omega Bio Tek E.Z.N.A.组织DNA KIT D3396-01 OMEGA BIO TEK MAG-BIND®血液和组织DNA HDQ 96 KIT M6399-00 OMAMGA BIO TEK E.Z.N.A. Kit CMG-196 Promega Wizard Genomic DNA Purification Kit A1120 Promega Wizard SV Genomic DNA Purification System A2360 Promega MagaZorb DNA Mini-Prep Kit MB1004 Promega ReliaPrep gDNA Tissue Miniprep System A2051 Promega Wizard HMW DNA Extraction Kit A2920 Promega ReliaPrep™ Blood gDNA Miniprep System A5081 Qiagen DNEasy Blood and Tissue Kit 69504 Qiagen QIAamp Fast DNA Tissue Kit 51404 Qiagen Puregene Tissue Kit 158063 Qiagen QIAGEN Genomic-tips 20/G 10223 Qiagen MagAttract HMW DNA Kit 67563 Qiagen QIAamp DNA Blood Mini Kit 51104 Qiagen Blood & Cell Culture DNA Mini Kit (25) 13323 Sigma-Aldrich Extract-N-AMP Tissue PCR Kit XNAT2-1KT Sigma-Aldrich GenElute Blood Genomic DNA Kit NA2010-1KT Takara NucleoSpin Tissue 740952.5 Takara NucleoMag Tissue 744300.1 Takara NucleoSpin® 96 DNA RapidLyse 740110.1 Takara NucleoBond HMW DNA 740160.2 Takara NucleoMag DNA Swab 744601.1 Takara NucleoSpin® Blood 740951.5 Takara NucleoMag® Blood 200 µL 744501.1 Thermo DNA Extract All Reagents Kit 4403319 Thermo MagMax DNA Multi-Sample Kit 4413020 Thermo JetFlex Genomic DNA Purification Kit A30700 Thermo GeneJet Genomic DNA Purification Kit K0721 Thermo GeneJet Genomic DNA Purification Kit K0721 Thermo PureLink Genomic DNA Mini Kit K182001 Thermo PureLink Genomic DNA Mini Kit K182001 Thermo ChargeSwitch gDNA Mini Tissue Kit CS11204 Zymo Quick-DNA MiniPrep D3024 Zymo Quick-DNA Miniprep Plus D4068 Zymo Quick-DNA magbead Plus Kit D4081 Zymo Quick-DNA HMW HMW Magbead d6060
eppendorf®,Eppendorf品牌设计,Epmotion®和EppendorfThermomixer®是德国Eppendorf SE的注册商标。Quick-DNA™是美国Zymo Research Corporation的注册商标。保留所有权利,包括图形和图像。版权所有©2024 Eppendorf SE
在37°C,200 rpm时10分钟。RNA,并在冰上被5 µL冰冷1 N NaOH碎片碎片10分钟。用25 µl 1 M Tris-HCl(pH 6.8)中和后,将RNA再次用异丙醇沉淀。Biotin RNA被预洗的dynabeads™M-280链霉亲和蛋白珠(Invitrogen,#11206D)富含,被高盐缓冲液,分别降水缓冲液,低盐缓冲液洗涤,然后用Trizol和Zymo rna Clean&Compentor(Zymoresearch(Zymoresearch)(Zymoresearch)在珠子上提取。用20°C的100 µM VRA3 RNA适配器与1 µL的100 µM VRA 3 RNA连接过夜。然后将生物素RNA富集并再次提取。RNA通过RPPH(NEB,#M0356)在5'端修饰,并通过T4 PNK(NEB,#M0201L)进行了羟基修复。被Zymo RNA清洁和浓缩器提取后
1尼日利亚纽约州尼日利亚大学尼日利亚大学植物科学与生物技术系。2 Inqaba Biotec West Africa Ltd.,尼日利亚伊巴丹Moniya的IITA巴士站对面。通讯作者:isuosuo chinyere chioma。由于成本,速度和安全性,可以使用几种商用套件代替常规提取方法。因此,使用Zymo Research Mini植物/种子DNA提取试剂盒来确定86种非洲treculia Africana品种的DNA提取物的产量和纯度。Nanodrop Thermo Scientific™Nanodrop™一种微型紫外线分光光度计用于确定提取的DNA的质量和数量。使用内部转录的间隔1和2(其1和2)通过聚合酶链反应扩增提取的DNA。在1%琼脂糖凝胶电泳上运行扩增子。记录了A260/280和A260/230的浓度(NG/µL)和样品的A260/230。配件之间的浓度和纯度值各不相同。A260/280的纯度分别在1.5到2.18之间,分别从B22和B43获得,而A260/230的比例分别为0.93至39.56,分别从CANSKIONS C46 - AB6获得。在A260/280比率和A260/230比率下得出的值主要在1.8 - 2.0的可接受范围内,这表明ZR试剂盒可以消除污染物。因此,可以方便地进行下游应用,例如PCR和测序。在琼脂糖凝胶图像上揭示了使用Zymo Research Mini Prep作为DNA提取试剂盒的适用性和效率。Zymo Research DNA提取试剂盒适合于从treculia物种的种子中提取DNA。
液氮 n/an/a 关键商业检测 NEBuilder® HiFi DNA 组装预混液 New England Biolabs E2621S BsaIHF®v2 (20 U/µL) New England Biolabs R3733S Phusion TM 高保真 DNA 聚合酶 Thermo Fisher Scientific F530S MluI (10 U/µL) Thermo Fisher Scientific ER0561 ApaI (10 U/µL) Thermo Fisher Scientific ER1411 XhoI (10 U/µL) Thermo Fisher Scientific ER0691 EcoRI (10 U/µL) Thermo Fisher Scientific ER0271 RevertAid TM 逆转录酶 Thermo Fisher Scientific EP0441 RiboLock RNase 抑制剂 (40 U/μL) Thermo Fisher Scientific EO0381 NucleoSpin® 质粒试剂盒 Macherey-Nagel 740588.250 Zymoclean 凝胶 DNA 回收试剂盒 Zymo Research D4001 Zymo-Spin I Zymo Research C1003-250 嗜热菌 (Tth) DNA 聚合酶 Biotools 10.003 寡核苷酸 D1789 GGGAATCAATCACAGTGTTGGC
图 1. 比较类似产品配置中 DH5α 感受态大肠杆菌与 NEB ™ 5-alpha 和 Zymo ™ Mix and Go! ™ 5α 感受态细胞的转化效率。根据制造商推荐的方案,使用 10 pg pUC19 DNA 进行三次转化来测量转化效率。每次转化均进行两次接种。