Jul 1, 2022 · 滚环扩增(rca)-fish的原理是先杂交并将挂锁探针连接到特定的mrna靶点，然后扩增挂锁探针。 在smFISH和RCA-FISH的基础上，为了进一步提高检测效率，提高亮度，降低总体成本。

RCA是以一小段环状寡核苷酸为模板，以dNTPs为原料，在DNA/RNA聚合酶作用下扩增产生一条长重复单链DNA/RNA，如下图所示。 滚环扩增方法目前已经广泛应用于 基因组学 、 蛋白组学 、 分子诊断 、 生物传感 、 药物研发 等领域 [2]。 RCA工作原理. （1）滚环扩增的方法模板必须是环状的，如果针对线性基因进行扩增，就需要 锁环探针 （padlock probe），锁环探针的两端具有与靶基因的互补序列，通过锁环探针识别靶基因并结合形成不完全闭合环状寡核苷酸，之后在 …

Jun 13, 2022 · RCA-FISH, clampFISH, HCR-FISH, RNAscope and SABER-FISH enhance signals by amplifying the primary probes that hybridize to the RNA target by RCA, by secondary and tertiary probes, or by...

滚环扩增(rca)-fish的原理是先杂交并将挂锁探针连接到特定的mrna靶点，然后扩增挂锁探针。 在smFISH和RCA-FISH的基础上，为了进一步提高检测效率，提高亮度，降低总体成本。

Sep 30, 2019 · 传统的荧光原位杂交（FISH）需要DNA变性，同时在体外与sgRNA组装在一起的荧光标记dCas9（CASFISH）仅检测固定样本中的基因组位点，这就限制了实时追踪。 在一项新的研究中，来自美国斯坦福大学、卡斯迪加学校和中国浙江大学的研究人员报道了一种称为CRISPR活细胞荧光原位杂交（CRISPR live-cell fluorescent in situ hybridization, CRISPR LiveFISH）的实时成像方法，从而允许研究活细胞中的各种染色体功能。 相关研究结果发表 …

Nov 28, 2018 · Using this combined RCA-smFISH (RollFISH) approach, we visualize and accurately quantify clinically relevant biomarkers, at single-cell resolution, across entire FFPE tissue sections. We conclude...

Jun 15, 2020 · FISH using rolling circle amplification (RCA), which uses complementary DNA (cDNA) generated from mRNA by RT, has a potential to detect a single mRNA molecule in a...

Dec 1, 2019 · Rolling circle amplification (RCA) is an isothermal nucleic acid amplification technique where a short DNA/RNA primer is amplified using a circular DNA template and special DNA/RNA polymerases (Phi29, Bst, and Vent (exo-) DNA polymerase for DNA, and T7 RNA polymerase for RNA) to form a long concatemer single-stranded DNA/RNA that contains hundre...

RCA-FISH (rolling circle amplification-Fluorescence in Situ Hybridisation) RCA-FISH is based on the in situ reverse-transcription of mRNA, a subsequent signal amplification using a loop-shaped DNA target probe as starting point for the RCA amplification and followed by FISH detection of the so amplified DNA (Table 1 ) 12 .

As an alternative to smFISH, in situ rolling circle amplification (RCA) is a method that may potentially achieve highly specific imaging of mRNA in single cells with near-single-molecule resolution. RCA can achieve localized isothermal amplification to provide information about the localization of target molecules.