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 ...
使用 Single Cell-to-CT™ 试剂盒检测单细胞当量表明,与具有出色技术重现性的100细胞样品对照品相比,qPCR 结果具有适当的线性和灵敏度(如图所示)。与预期一致,单个细胞因其固有的生物异质性而表现出了更大的变异性。此外,通过包含 cDNA 预扩增步骤,在实时荧光定量 PCR 之前准确扩增目标物。该关键步骤以无偏倚的...
(2005) Quantitative single-cell RT-PCR and Ca 2+ imaging in brain slices. Pflugers Arch 451 , 716–726.Durand G, Marandi N, Herberger S, Blum R, Konnerth A: Quantitative single-cell RT-PCR and Ca 2+ imaging in brain slices. Pflugers Arch 2006, 451: 716–726....
(1) The entire cell is harvested and reverse transcribed. (2) Mutant sequences, included as internal controls, explicitly monitor the efficiency of RT and PCR in each reaction tube. (3) Multiple targets are independently amplified from each cell, including a constitutively expressed housekeeping ...
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...
R. Genome-wide single-cell analysis of recombination activity and de novo mutation rates in human sperm. Cell 150, 402–412 (2012). Article PubMed PubMed Central CAS Google Scholar White, A. et al. High-throughput microfluidic single-cell RT-qPCR. Proc. Natl Acad. Sci. USA 108, 13999...
随后通过RT-PCR、巢式PCR得到抗原特异性单个B细胞抗体可变区基因。 3. 表达、筛选和鉴定抗原特异性抗体 FACS分选的单B细胞抗体基因扩增、载体构建、抗体表达及活性验证是必要步骤。表达系统常用的是原核表达系统(例如大肠杆菌)或真核表达系统。在大肠杆菌中,通常表达抗体的抗原结合片段(Fab),而在哺乳动物中,可以表达...
Single Cell Sequence Specific Amplification Kit P621-01 Version 5.1 Vazyme biotech co., ltd. 产品简介 Single Cell Squence Specific Amplification Kit 是基于一步RT-PCR的扩增方法,用于实现单细胞或者微量总RNA中转录组的扩增,便于您 揭开单个细胞之间不同基因的表达水平。本试剂盒实现了RNA提取纯化、逆转录和...
单细胞全转录组扩增试剂盒(Single Cell WTA Kit)提供了更简单、更快速和更高质量的方法,包括质粒提取、胶回收、PCR产物纯化、基因组提取、RNA分离纯化、PCR/RT-PCR、荧光定量、克隆载体和感受态细胞制备等数百种实验常用试剂盒。 单细胞全转录组扩增试剂盒(Single Cell WTA Kit) 产品及特点 本产品是用于对从单个...
Detection of Serum HDV RNA by RT-PCR 2004, Springer Protocols less References Jorgensen JL, Esser U, de St F, Groth B, Reay PA, Davis MM (1992) Mapping T-cell receptor-peptide contacts by variant peptide immunization of single-chain transgenics. Nature 355:224–230 Davis MM, Bjorkma...