Total RNA (1 µg) was reverse-transcribed into cDNA using the cDNA synthesis kit (Takara Bio Inc., Shiga, Japan) according to the manufacturer’s instructions. Briefly, RNA was mixed with a mixture of Oligo DT p...
DNA polymerase activity with Poly (A)·oligo(dT) as a template‐primer has been found in preimplantation mouse embryos. A sharp increase in enzyme activity was observed at the time of formation of inner cell mass (ICM) and trophoblast cells. Differential inhibition of enzyme activity before and...
For example, adding Poly (A) to miRNA provides an oligo-dT primer binding site for cDNA synthesis. 3. Improve the translation efficiency of RNA in eukaryotic cells. Preservation system 20mM Tris-HCl; 300mM NaCl; 1mM DTT; 1mM EDTA; 0.1% TritonX-100; 50% (v/v) Glycerol, pH 7.5. ...
MicroRNA (miRNA)是一类长度为19-23nt的内源性非编码单链小分子RNA,是一种重要的基因表达调控因子。本试剂盒采用在miRNA 3’末端加多聚A尾Poly(A),再使用Oligo(dT)-universal tag通用逆转录引物进行逆转录反应,生成miRNA对应的cDNA第一链,最后采用miRNA的特异性正向引物和通用反向引物进行特异性的miRNA qPCR检测...
(polyA/RT)和 qPCR 定量检测三个步骤(见图 1).本方法使用由 5~7 个特异碱基和 11 个 dT 组成的 S-Poly(T)Plus 引物对 poly(A) tailed miRNA 进行特异性逆转 录(RT),显著增强了 poly(A) tailed miRNA 与特异 RT 引物之间的结合力,从而大幅度提高 cDNA 的合成效率及其特异性,最终使后续 qPCR 检测的...
DNA polymerase activity with Poly (A)·oligo(dT) as a template-primer has been found in preimplantation mouse embryos. A sharp increase in enzyme activity was observed at the time of formation of inner cell mass (ICM) and trophoblast cells. Differential inhibition of enzyme activity before and...
Krug, M. S. & Berger, S. L. First-strand cDNA synthesis primed with oligo(dT).Methods Enzymol.152,316–25 (1987). Winter, G. & Brownlee, G. G. 3'End labelling of RNA with 32P suitable for rapid gel sequencing.Nucleic Acids Res.5,3129–39 (1978). ...
First, 18S has no poly(A) tail, so it is commonly reverse transcribed with specific primers or random hexamers, independently from poly(dT)-primed transcripts. Secondly, due to its abundance, the 18S cDNA must be extensively diluted to be comparable to the tested genes. In this study, 18S...
We have analyzed a systematic flaw in the current system of gene identification: the oligo(dT) primer widely used for cDNA synthesis generates a high frequency of truncated cDNAs through internal poly(A) priming. Such truncated cDNAs may contribute to 12% of the expressed sequence tags in the ...
? Flexible: Use polyadenylated RNAs in a variety of applications such as stabilizing in vitro transcribed RNA and adding primer (oligo-dT) binding sites to any RNA for first-strand cDNA synthesis ? Fast and Easy: Simple protocol produces polyadenylated RNA molecules quickly ...