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...
(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...
The RT primer contains six miRNA-specific bases and a oligo(dT) sequence. The amplification products are detected by a universal Taqman probe. In general, the miRNAs are quantified by measuring the temperature cycles to cross a fluorescence threshold (Ct values) and only provide a relative ...
a Streptavidin pull-down assays of the binding of biotinylated poly(dA:dT) (Bio-poly(dA:dT)), biotinylated long poly(I:C) (Bio-poly(I:C)), or biotinylated reovirus dsRNA (Bio-ReoRNA) to purified HA-tagged PARP9 or PARP13 in HEK 293T cells. b Streptavidin pull-down assays of the...
dT primer (5′-AAGCAGTGGTATCAACGCAGAGTACT30VN-3′, where “N” is A, T, C, or G and “V” is A, C, or G) for reverse transcription to construct the complementary DNA (cDNA) library. The two terminal nucleotides “N” and “V” anchor the reverse transcriptase (RT) primer to...
A common approach to the synthesis of cDNA for RT-PCR analysis or the synthesis of cDNA libraries begins with the selection of poly(A)+ mRNA. To initiate conversion of mRNA into first-strand cDNA, an oligo(dT) primer is annealed to the mRNA template to provide the requisite 3′-OH group...
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 ...
Polycarbamate Standard 代森福美锌标准品 品牌:FUJIFILM Wako CAS No.:64440-88-6 储存条件:2-10℃ 纯度:95.0 – 105.0% (Titration) 产品编号 (生产商编号) 等级规格运输包装零售价(RMB)库存情况参考值 167-26581 for Pesticide Residue Analysis100mg–1,010.00–– ...
Angiogenic marker (hypoxia-inducible factor 1-alpha (HIF-1α) and VEGF and inflammatory gene marker expression (tumor necrosis factor-α (TNF-α), IL-1β, IL-6, IL-4, IL-10, and Arg-1) were quantified using RT-PCR. β-actin was the housekeeping gene. The primer sequences are ...
RNA was analyzed by RT-PCR with oligo(dT) or random hexamer primers, and the amplified products were sequenced. Five of the mutations, FS211X (type 1), FS288X, FS546X, FS588X, and FS737X, truncate the protein in the α-subunit, yielding truncated α-subunit and no β-subunit. ...