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一般描述
GenElute mRNA小量制备试剂盒可以简单、方便地从预先分离的总RNA中纯化多腺苷酸化mRNA。在10分钟的孵育时间内，Oligo(dT)聚苯乙烯微珠与poly(A)+ mRNA结合在一起。在微量离心滤器中洗涤以去除污染物之后，在100 mL缓冲液中洗脱poly(A)+ mRNA。在40分钟内，即可从总RNA中纯化出mRNA。
纯化所得的mRNA可直接用于Northern分析、反转录、PCR、阵列标记及其他常规应用。
纯化所得的mRNA可直接用于Northern分析、反转录、PCR、阵列标记及其他常规应用。
适于cDNA合成、表达谱及其他需要从超高丰度的rRNA和tRNA中分离mRNA的操作。GenElute mRNA试剂盒为从之前制备的总RNA中分离多腺苷酸化mRNA，或直接从哺乳动物细胞和组织中分离多腺苷酸化mRNA提供了方便的操作方法。直接制备mRNA时，需采用SDS/蛋白酶K降解法破碎细胞或组织，从而释放出RNA并去除RNases。这两种试剂盒均采用与1 μm聚苯乙烯微珠共价偶联的oligo dT30，通过杂交法捕获多腺苷酸化的mRNA。在杂交过程中，聚苯乙烯微珠继续保持悬浮状态，无需进行混合或振荡（这在纤维素或磁微粒方法中较为常见）。oligo(dT) 聚苯乙烯微珠（2-3步）可以采用比常用的oligo(dT) 纤维素微粒法（至少10步）更少的严格洗涤步骤，获得更纯净的mRNA，聚苯乙烯的种类是可选的。采用GenElute试剂盒时，基于mRNA的复合物在一个微量离心滤器上洗涤，然后在10 mM Tris-HCL, pH 7.5.缓冲液洗脱。通过上述任一种试剂盒制备的mRNA均适用于Northern杂交、表达阵列或芯片杂交、cDNA合成、文库构建等广泛的下游应用。
应用
纯化所得的mRNA可直接用于Northern分析、反转录、PCR、阵列标记及其他常规应用。
特点和优势
- oligo(dT) 聚苯乙烯微珠可在10分钟内捕获mRNA
- Oligo(dT) 聚苯乙烯微珠所需的洗涤步骤更少
- 可在40分钟内从之前纯化的总RNA中分离出Poly (A)+ mRNA
- 同时分离RNA、DNA和蛋白质的快速、方便的试剂
- 适合于各种数量规模的组织或细胞，可同时提取多份样本
- 是最高效的总RNA分离方法之一。以新鲜组织或细胞为起始材料，仅需1小时，即可完成纯化
其他说明
更多信息，请见www.sigma-aldrich.com/mrna。
法律信息
GenElute is a trademark of Sigma-Aldrich Co. LLC
储存分类代码
10 - Combustible liquids
Caitlin M A Simopoulos et al.
G3 (Bethesda, Md.), 9(8), 2511-2520 (2019-06-27)
Long non-coding RNAs (lncRNAs) represent a diverse class of regulatory loci with roles in development and stress responses throughout all kingdoms of life. LncRNAs, however, remain under-studied in plants compared to animal systems. To address this deficiency, we applied a
Liping Wu et al.
Frontiers in immunology, 13, 839677-839677 (2022-06-28)
Host translation is generally modulated by viral infection, including duck hepatitis A virus (DHAV) infection. Previously, we reported that cellular protein synthesis in a cell model of duck embryo fibroblasts is significantly inhibited by DHAV infection but not viral proteins
Gang Luo et al.
Foods (Basel, Switzerland), 11(11) (2022-06-11)
N6-methyladenosine (m6A) is the most prevalent internal mRNA modification in eukaryotes. The M6A modification plays an important role in transcription and cell function. The mechanism by which m6A modification regulates meat quality remains elusive. In this study, gene knockout and
Jianzi Lan et al.
Cellular & molecular biology letters, 27(1), 51-51 (2022-06-28)
Diabetic nephropathy (DN) is prevalent in patients with diabetes. N6-methyladenosine (m6A) methylation has been found to cause modification of nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing (NLRP) 3, which is involved in cell pyroptosis and inflammation. WTAP is a
Kaiping Deng et al.
Molecular therapy. Nucleic acids, 26, 34-48 (2021-09-14)
N6-methyladenosine (m6A) modification plays a critical role in mammalian development. However, the role of m6A in the skeletal muscle development remains largely unknown. Here, we report a global m6A modification pattern of goat skeletal muscle at two key development stages
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