We modified the single cell RT-PCR approach previously described (2) into a reproducible and friendly technique, for studying and characterization of certain biochemical events in mast cells cocultured with fibroblasts (3). This original work clearly showed enhancement of interleukin (IL)-3 ...
For Use With (Application)RT-PCR No. of Reactions50 Reactions Sample TypeMammalian Cells Starting Material1-10 Cells Unit SizeEach Contents & Storage 50 μL Single Cell DNase I, 0.5 mL Single Cell Lysis Solution (store at 4°C), 50 μL Single Cell Stop Solution, 75 μL Single Cell Sup...
方法:采用倍比稀释相对定量单细胞RT-PCR方法分离纯化人外周血中性粒细胞并提取总RNA,针对IL-8RβmRNA设计引物,进行RT-PCR确认IL-8Rβ在中性粒细胞中的基因表达并优化反应条件。 2) Single cell nested RT-PCR 单细胞巢式RT-PCR 3) in-cell reverse transcription poly-merase chain reaction(in-cellRT-PCR) ...
由于单个细胞中的mRNA数量很少,大约只有1 pg左右,因而与常规的RT-PCR相比,单细胞RT-PCR在各个环节上都有着显著差异。常规RT-PCR首先要将组织中的mRNA或总RNA抽提出来之后才能进行后续步骤,单细胞RT-PCR显然无法进行抽提步骤,因而在获取单个细胞后,在适当体积的裂解液中将细胞裂解,然后直接以裂解产物为模板进行RT-PCR...
Durand, G.M. et al. (2006) Quantitative single-cell RT-PCR and Ca2+ imaging in brain slices. Pflugers Arch. 451, 716-726Durand G, Marandi N, Herberger S, Blum R, Konnerth A: Quantitative single-cell RT-PCR and Ca2+ imaging in brain slices. Pflugers Arch 2006, 451:716-726....
由于单个细胞中的mRNA数量很少,大约只有1 pg左右,因而与常规的RT-PCR相比,单细胞RT-PCR在各个环节上都有着显著差异。常规RT-PCR首先要将组织中的mRNA或总RNA抽提出来之后才能进行后续步骤,单细胞RT-PCR显然无法进行抽提步骤,因而在获取单个细胞后,在适当体积的裂解液中将细胞裂解,然后直接以裂解产物为模板进行RT-PCR...
•单细胞实时荧光定量 RT-PCR 的预优化工作流程•单细胞分析的较高灵敏度•性能和重现性优于替代方法•全套试剂盒方便且易于执行方案预优化方案可确保成功并节省时间每个工作流程均从简单的7分钟样品制备开始,其中以极少的工艺将细胞有效、可重现地裂解为与下游 RT-PCR 兼容的溶液,无需纯化。随后使用 ...
随后同时对每个单个细胞的TCRαβ转录本和DNA barcode进行RT-PCR扩增,细胞barcode可用于在单细胞水平将TCR序列与其同源抗原进行配对。Zhang等采用此方法同时筛选出超过150中癌新抗原和野生型多肽对,这说明可以利用TetTCR-seq高通量识别具有功能活性的新抗原特异性TCR。
In this study we illustrate the use of a very sensitive protocol of semiquantitative single-cell RT-PCR that we have developed. This technique allows the simultaneous analysis of up to eight genes on a single cell. Here we show the experiments performed to validate this novel assay. As an ...
et al. Allele-specific expression patterns of interleukin-2 and Pax-5 revealed by a sensitive single-cell RT-PCR analysis. Curr. Biol. 10, 789–792 (2000). CAS PubMed Google Scholar Esumi, S. et al. Monoallelic yet combinatorial expression of variable exons of the protocadherin-α gene...