从植物和真菌中纯化总RNA

  • 30分钟即可获得高品质的总RNA
  • 无需酚/氯仿抽提
  • 无需CsCl梯度离心,无需LiCl或乙醇沉淀
  • 出色的RNA回收率
  • 即用型RNA适合多种下游应用

RNeasy Plant Mini Kit含QIAshredder离心柱和RNeasy离心柱,前者用于匀质化和过滤高粘度的植物或真菌的裂解物,后者使用硅胶膜技术纯化至多100 μg高品质RNA。纯化过程可在QIAcube全自动核酸纯化仪上自动化进行。该试剂盒也可配合TissueRuptor或TissueLyser体系使用,高效破碎并匀质化植物样本。

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RNeasy Plant Mini Kit (20)
20 RNeasy Mini Spin Columns, 20 QIAshredder Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
74903
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RNeasy Plant Mini Kit (50)
50 RNeasy Mini Spin Columns, 50 QIAshredder Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
74904
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The RNeasy Plant Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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RNeasy Plant Mini procedure.|Detection of viruses.|Tissue specificity of histone H4 expression.|
|Total RNA was isolated from budwood obtained from infected apple (1), cherry (2), pear (3), and grapevine (4–8), leaf tissue from black currant (9, 10), and dormant tuber tissue from infected potato (11, 12) using a modification of the RNeasy protocol. Viral RNA was amplified using a one-tube RT-PCR. Amplification products were analyzed on a 1.5% agarose gel. M: DNA Markers VI, Boehringer Mannheim. (Data kindly provided by D.J. MacKenzie, Centre for Plant Health, Agriculture and Agri-Food Canada and M.A. McLean, Euro Nursery & Vineyard (West) Inc., Sidney, Canada.)|Formaldehyde agarose gel and northern blot of total RNA (7 µg) from indicated tissues of tomato. Blot was hybridized with a 32P-labeled DNA probe for the tomato histone H4 gene. M: 0.24-9.5 kb RNA ladder. (Plant tissues and histone H4 cDNA clone kindly provided by K. Theres, MPI for Breeding Research, Cologne, Germany.)|
Performance

RNeasy Plant Mini Kit适用于从各种植物和真菌样本中纯化总RNA,样本为10–100 mg的组织或100–1 x 107个细胞(参见"RNeasy Plant Mini Kit处理的样本")。离心柱的结合能力达100 µg RNA。100 mg植物组织的常规产量是25–65 µg RNA,但样本不同的发育阶段和生长条件可能导致不同的RNA产量(参见"100 mg组织的产量")。

RNeasy Plant Mini Kit 处理的样本
植物 丝状真菌
Anemone sp.
Arabidopsis thaliana1
Begonia sp.
Beta vulgaris (甜菜)2
Chlamydomonas reinhardii (单细胞藻类)
Chrysanthemum
Clarkia spp.3
Daucus carota (胡萝卜t)4
Diascia sp.
Dendranthema sp.
Euglena gracilis (单细胞藻类)
Funaria hygrometrica (苔)
Hordeum vulgare (大麦)
Lycopersicon esculentum (西红柿)5
Malus sp. (苹果)6
Nicotiana tabacum (烟草)
Oryza sativa (米)
Pelargonium sp. (天竺葵)
Petroselinum crispum (香菜)7
Pisum sativum (豌豆)
Prunus sp. (樱桃)6
Ranunculus sp.
Ribes nigrum (红醋栗)
Riccia fluitans (苔)
Sinapis arvensis (芥末)
Solanum tuberosum (土豆)8
Spinacia oleracea (菠菜)
Surfinia sp.
Triticum aestivum (小麦)
Vitis sp. (葡萄藤)6
Zea mays (玉米)
Acremonium crysogenum9
Fusarium avenaceum9
Young leaves or needles, except as indicated: 1 Flowers, buds, siliques, roots; 2 roots; 3 petals, stamen, stigma, sepals, style; 4 cultured cells; 5 mature flower, expanded leaves, shoot tip, stem, green fruit; 6 budwood; 7 seedlings; 8 dormant tuber; 9 mycelium
100 mg组织的产量
样本类型 产量
拟南芥叶 35 µg
玉米叶 25 µg
番茄叶 65 µg
烟草叶 60 µg

纯化的植物RNA,包含病毒RNA,洗脱体积30–100 µl(参见"Detection of viruses")。多聚糖等污染物被除去,洗脱得到的RNA适用于多种下游应用(参见"Tissue specificity of histone H4 expression")。

Principle

RNeasy Plant Mini Kit提供QIAshredder离心柱用于匀质化和通过离心法过滤高粘度的植物或真菌裂解物,同时采用 RNeasy Mini离心柱对RNA进行纯化。RNeasy纯化技术整合了异硫氰酸胍裂解的严谨性和硅胶膜纯化的速度和纯度,简化了总RNA纯化技术。RNeasy试剂盒提供高品质的RNA,DNA污染达到最低水平。对微量DNA非常敏感的RNA应用,可在RNeasy操作流程中进行方便的柱上DNase处理,去除残留的DNA。

Procedure
样本首先被裂解,并在QIAshredder离心柱中匀质化。在裂解液中加入乙醇以提供合适的结合条件。将裂解液上样到RNeasy硅胶膜上,RNA与硅胶膜结合,各种污染物被高效洗去。浓缩的纯RNA被水洗脱(参见"RNeasy Plant Mini procedure")。
Applications

RNeasy技术纯化得到的高品质RNA适合各种下游应用,包括:

Northern印迹、点印迹和狭缝印迹
终点式RT-PCR
定量real-time RT-PCR
芯片分析
Poly A+ RNA筛选
Feature
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 30–100 µl
Format Spin column
Main sample type Plant samples
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 10–100 mg
Technology Silica technology
Time per run or per prep 30 minutes
Yield 25–60 µg

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Kit Handbooks
1
RNeasy Mini Kit - For purification of total RNA from animal cells, animal tissues, bacteria, and yeast, and for RNA cleanup; RNeasy Protect Mini Kit - For immediate stabilization of RNA in harvested animal tissues and subsequent total RNA purification; RNeasy Plant Mini Kit - For purification of total RNA from plants and filamentous fungi
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