一步法miRNA反转录试剂盒
由于miRNA(microRNA)分子序列较小,仅20nt左右,没有真核生物特有的Poly(A)尾巴结构,采用传统Oligo dT引物和随机引物很难获得从miRNA到cDNA的反转录产物。目前普遍采用的解决方案有两种:特异性反转录引物(图1A所示)、两步加A法反转录方案(图1B所示)。 HaiGene公司开发出一套全新的一步法miRNA反转录试剂盒,基于Peter改良的miR-Q技术,可以对成熟的miRNA同时完成Poly(A)反应和反转录反应,直接获得含有特异性tag和15(T)的cDNA产物(如图2所示)。该方法使得miRNA反转录与mRNA的反转录一样轻松,直接将提取的microRNA与反应缓冲液混合物混合并置于PCR仪上1小时即可获得高浓度的、包含所有miRNA分子的cDNA产物,获得的cDNA产物适合于采用SYBR Green染料法进行RealTime PCR进行定量检测。 | ||
组分 | ||
组分名称数量4XOne step miRNA RT Solution125 μl/500 μl10×miRNA RT Primer50 μl/200 μlRNase Free H2O1 ml/1 ml×2 | ||
注:如4XOne step miRNA RT Solution出现沉淀属正常现象,可用手捂化,混合均匀后使用不影响实验结果。 | ||
一步法microRNA反转录原理 | ||
储存 | ||
请置于-20°C,可保存2年;避免反复冻融。 | ||
应用本产品文献 | ||
1.MicroRNA‐365 modulates astrocyte conversion into neuron in adult rat brain after stroke by targeting Pax6. 2.Grape Seed Procyanidin B2 Protects Porcine Ovarian Granulosa Cells against Oxidative Stress-Induced Apoptosis by Upregulating let-7a Expression. 3.Exosomes Derived From MicroRNA-148b-3p-Overexpressing Human Umbilical Cord Mesenchymal Stem Cells Restrain Breast Cancer Progression. 4.MicroRNA Expression Profiling in Newcastle Disease Virus-Infected DF-1 Cells by Deep Sequencing. 5.SLC34A2 promotes neuroblastoma cell stemness via enhancement of miR‐25/Gsk3β‐mediated activation of Wnt/β‐catenin signaling. 6.LncRNA SNHG16 Functions as an Oncogene by Sponging MiR-135a and Promotes JAK2/STAT3 Signal Pathway in Gastric Cancer. 7. Circular RNA profiling identified an abundant circular RNA circTMTC1 that inhibits chicken skeletal muscle satellite cell differentiation by sponging miR-128-3p. 8.An innovative “unlocked mechanism” by a double key avenue for one-pot detection of microRNA-21 and microRNA-141. 9.Upregulation of miR-375 level ameliorates morphine analgesic tolerance in mouse dorsal root ganglia by inhibiting the JAK2/STAT3 pathway. 10.Transcriptome-Wide Identification and Characterization of MicroRNAs from Castor Bean (Ricinus communis L.). |