Monarch 总 RNA 小量提取试剂盒（ #T2010） |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

Monarch 总 RNA 小量提取试剂盒（ #T2010） |

Monarch 总 RNA 小量提取试剂盒是一款可用于样本保存、细胞裂解、gDNA 去除，以及提取纯化总 RNA 的综合性解决方案试剂盒，适用于多种生物样本，如培养细胞、血液和哺乳动物组织。此外，也适用于难裂解样本，如细菌、酵母和植物，只要增加一步来提高裂解效率。该试剂盒还可用于纯化酶促反应后的样本，或者使用 TRIzol® 提取的样本。纯化后的 RNA 具有极高质量：A260/280 和 A260/230 比值均 > 1.8，高 RIN 值和无 gDNA 残留；并且提取的 RNA 片段包含全长 rRNA 和完整的 miRNA。此外，改变结合条件可以选择性地提取或去除小于 200 nt 的 RNA，包含 miRNA、5S rRNA 和 tRNA。纯化后的 RNA 可用于 RT-qPCR、cDNA 合成、RNA-seq、Northern blot 分析等下游实验。

Monarch 总 RNA 小量提取试剂盒实验流程

Monarch 总 RNA 小量提取试剂盒（ #T2010） |

 

规格：

结合能力： 100 µg RNA
RNA 大小： ≥ 20 nt
纯度： A260/280（比值 ≥ 1.8），A260/230（比值 ≥ 1.8）
组织起始样本量： 高产量组织最多 20 mg
  低产量组织最多 50 mg
培养的哺乳动物细胞和血细胞（外周血单核细胞、白细胞）起始样本量： 最多 1 x 107
哺乳动物全血起始样本量： 最多 3 ml
细菌起始样本量： 最多 1 x 109
酵母起始样本量： 最多 5 x 107
植物起始样本量： 最多 100 mg
洗脱体积： 30 – 100 μl
产量： 取决于样本类型
兼容的下游实验： NGS RNA 文库制备、RT-PCR、RT-qPCR 和 Northern blot 分析

 

经过验证的样本类型：

  • HeLa 细胞
  • HEK 293 细胞
  • NIH 3T3 细胞
  • 人血
  • 大鼠血
  • 外周血单核细胞
  • 血浆
  • 血清
  • 有核血
  • 大鼠肝
  • 大鼠脾
  • 大鼠肾
  • 大鼠脑
  • 大鼠肌肉
  • 小鼠肌肉
  • 小鼠心脏
  • 小鼠肾
  • 斑马鱼幼体
  • 果蝇
  • 酿酒酵母
  • 大肠杆菌
  • 蜡样芽孢杆菌
  • 番茄叶
  • 玉米叶
  • 唾液
  • 口腔细胞

 

数据图：

图 1： Monarch 纯化后的 RNA 具有高质量，可用于多种下游实验

Monarch 总 RNA 小量提取试剂盒（ #T2010） |
使用 Monarch 总 RNA 小量提取试剂盒（NEB #T2010）从多种样本类型中纯化总 RNA。取适量提取的 RNA 样本，使用 Nano 或 Pico 6000 RNA 芯片，在 Agilent® Bioanalyzer® 2100 上检测（酿酒酵母 RNA 检测使用植物 Nano 分析模式）。通过 RIN 值和 O.D. 值分析 RNA 的完整性和纯度。随后验证与下游应用的兼容性：（A）RT-PCR，使用 ProtoScript II 反转录酶（NEB #M0368）/LongAmp Taq DNA 聚合酶（NEB #M0323）分析；（B）NGS 文库制备，使用 NEBNext® Ultra™ II RNA 文库制备试剂盒（NEB #E7760）分析；和（C）RT-qPCR，使用 Luna 通用一步法 RT- qPCR 试剂盒（NEB #E3005）分析。

 

图 2： Monarch 总 RNA 小量提取试剂盒可以从少至 100 个 Hela 细胞中提取高质量 RNA

Monarch 总 RNA 小量提取试剂盒（ #T2010） |
使用 Monarch 总 RNA 小量提取试剂盒（NEB #T2010）从不同数量的 HeLa 细胞（跨越 5 个数量级）中分离总 RNA，并使用 100 µl 无核酸酶水洗脱。（A）使用 Bioanalyzer Pico 芯片分析样本，泳道下方显示 RIN 值和总产量。（B）电泳图作为参考。

 

图 3：Monarch 总 RNA 小量提取试剂盒成功纯化小于 200 nt 的小 RNA，能够更真实地代表总 RNA 库

Monarch 总 RNA 小量提取试剂盒（ #T2010） |
使用 Monarch 总 RNA 小量提取试剂盒（NEB #T2010）和 Qiagen 的 RNeasy 小量提取试剂盒，分别从 HEK293 细胞、小鼠心脏、大鼠脑或大鼠脾中提取纯化 RNA 。使用小 RNA 芯片在 Bioanalyzer 2100 上检测等量的样本。Monarch 纯化后的 RNA 包含更多小于 200 nt 的 RNA。

 

产品类别:
Total RNA Extraction & Purification Products,
RNA Extraction and Purification,
Nucleic Acid Purification Products

  • 试剂盒组成

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • T2010S     25    
        Monarch® gDNA Removal Columns T2017-1 25 1 x 50 columns Not Applicable
        Monarch® RNA Purification Columns T2007-1 25 1 x 50 columns Not Applicable
        Monarch® Spin Collection Tubes T2118-1 25 2 x 50 tubes Not Applicable
        Monarch® DNA/RNA Protection Reagent T2011-1 25 1 x 28 ml Not Applicable
        Monarch® RNA Lysis Buffer T2012-1 25 1 x 50 ml Not Applicable
        Monarch® Proteinase K Reaction Buffer T2003-1 25 1 x 4 ml Not Applicable
        Monarch® DNase I Reaction Buffer T2005-1 25 1 x 4 ml Not Applicable
        Monarch® RNA Priming Buffer T2013-1 25 1 x 28 ml Not Applicable
        Monarch® RNA Wash Buffer T2014-1 25 1 x 25 ml Not Applicable
        Monarch® Nuclease-free Water T2006-1 25 1 x 13 ml Not Applicable
        Monarch® Proteinase K T2001-1 25 1 x 20 mg 20 mg/ml
        Monarch® Proteinase K Resuspension Buffer T2002-1 25 1 x 1.5 ml Not Applicable
        Monarch® DNase I T2004-1 25 1 x 275 units 1 units/μl

  • 相关产品

    相关产品

    • Luna® 通用一步法 RT-qPCR 试剂盒
    • Luna® 通用探针一步法 RT-qPCR 试剂盒
    • e6560-protoscript-ii-first-strand-cdna-synthesis-kit
    • NEBNext® Ultra™ II RNA 非链特异性文库制备试剂盒
    • e7760-nebnext-ultra-ii-directional-rna-library-prep-kit-for-illumina
    • e7765-nebnext-ultra-ii-directional-rna-library-prep-with-sample-purification-beads
    • NEBNext® rRNA 去除试剂盒 – 人/小鼠/大鼠（含 RNA 纯化磁珠）
    • e7490-nebnext-polya-mrna-magnetic-isolation-module
    • m0368-protoscript-ii-reverse-transcriptase
    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）

    单独销售的组分

    • Nuclease-free Water
    • Monarch® DNA/RNA 保护试剂
    • Monarch® RNA 提取离心柱
    • Monarch® gDNA 去除离心柱
    • Monarch® RNA 裂解液
    • Monarch® RNA 初始洗涤缓冲液
    • t2014-monarch-rna-wash-buffer
    • Monarch® 总 RNA 小量提取酶套装
    • Monarch® 收集管 II

  • 注意事项

    1. 在复溶前，Monarch DNase I 和 Monarch 蛋白酶 K 可在室温下贮存。复溶后，请在 -20℃ 条件下贮存酶。建议分装 DNase I，以减少反复冻融次数。

操作说明、说明书 & 用法

  • 操作说明

    1. Quick Protocol for Total RNA Extraction (NEB #T2010)
    2. Download Quick Protocol Card T2010
    3. Total RNA Purification from Cultured Mammalian Cells using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    4. Total RNA Purification from Tissues and Leukocytes using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    5. Total RNA Purification from Mammalian Whole Blood Samples using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    6. Total RNA Purification from Tough-to-Lyse Samples (bacteria, yeast, plant, etc.) using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    7. FFPE Tissue Deparaffinization and Subsequent RNA Purification Using the Monarch Total RNA Miniprep Kit  (NEB #T2010)
    8. Total RNA Purification from TRIzol®-extracted Samples using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    9. Total RNA Purification from Samples in RNAlater® using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    10. Total RNA Purification from Samples Stored in Monarch DNA/RNA Protection Reagent using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    11. RNA Reaction Cleanup using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    12. In-tube DNase I Treatment after Purification with the Monarch Total RNA Miniprep Kit (NEB #T2010)
    13. Fractionation of Small and Large RNA Using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    14. RNA Extraction from Saliva, Buccal Swabs, and Nasopharyngeal Swabs using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    15. Automation of RNA Extraction from Saliva Using the Monarch Total RNA Miniprep Kit

  • 使用指南

    • General Guidelines for Successful RNA Purification Using the Monarch Total RNA Miniprep Kit
    • Guidance on Choosing Sample Input Amounts when Using the Monarch Total RNA Miniprep Kit
    • Guidelines for Purification of RNA from Cultured Mammalian Cells
    • Guidelines for RNA Purification from Bacteria
    • Guidelines for RNA Purification from Leukocytes (WBCs, PBMCs)
    • Guidelines for RNA Purification from Plants
    • Guidelines for RNA Purification from Tissues 
    • Guidelines for RNA Purification from Whole Blood
    • Guidelines for RNA Purification from Yeast
    • Guidelines for Sample Stabilization Using the Monarch DNA/RNA Protection Reagent

  • 应用实例

    • A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers
    • Purification of synthetic SARS-CoV-2 viral RNA from biological samples using the Monarch® Total RNA Miniprep Kit and the Monarch RNA Cleanup Kit

FAQs & 问题解决指南

  • FAQs

    1. Is the Monarch Total RNA Miniprep Kit compatible with Luna RT-qPCR Reagents?
    2. Is the Monarch Total RNA Miniprep Kit compatible with NEBNext reagents for RNA Library Prep?
    3. What is the lowest volume in which I can elute my RNA with the Monarch Total RNA Miniprep Kit (NEB #T2010)?
    4. How can I tell the difference between the gDNA Removal Column and the RNA Purification Column?
    5. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from yeast?
    6. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from bacteria?
    7. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from plants?
    8. How can I preserve my sample if I am not ready to extract RNA right after harvest?
    9. Can I purchase Monarch buffers and columns separately?
    10. I am seeing genomic DNA in my preps when using the Monarch Total RNA Miniprep Kit What could be the problem?
    11. Can I use the Monarch Total RNA Miniprep Kit for RNA cleanup after enzymatic manipulations?
    12. Can I skip the on-column DNase I treatment step of the Monarch Total RNA Miniprep Kit protocol?
    13. Can I skip the gDNA removal column step of the Monarch Total RNA Miniprep Kit protocol?
    14. Can I use leftover Proteinase K and DNase I from the Monarch Total RNA Miniprep Kit for other applications?
    15. Can I use NEB Proteinase K Molecular Biology Grade NEB P8107 in conjunction with the Monarch Total RNA Miniprep Kit?
    16. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from saliva?
    17. Can the Monarch Total RNA Miniprep Kit NEB T2010 be used to purify RNA from buccal cheek swabs?
    18. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from plasma or serum?
    19. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from nucleated blood (e.g. bird, fish, reptile)?
    20. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from drosophila?
    21. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from zebrafish embryos/larvae?
    22. Can the genomic DNA captured on the Monarch gDNA Removal Column (NEB #T2017) be eluted and used in downstream applications?
    23. Can I lyse my adherent cells directly in their wells when using the Monarch Total RNA Miniprep Kit?
    24. How can I assess RNA integrity and purity?
    25. Can I use NEB DNase I (#M0303) with the Monarch Total RNA Miniprep Kit?
    26. I would like to purchase the contents of the Monarch Total RNA Miniprep Kit (NEB #T2010S) separately. Which products do I need?
    27. Can extraction of RNA with the Monarch kits be automated?
    28. Is live SARS-CoV-2 virus inactivated during extraction with the Monarch Total RNA Miniprep Kit?

  • 问题解决指南

    • Troubleshooting Guide for Total RNA Extraction & Purification