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...
One strategy to achieve this goal combines whole cell electrophysiology and reverse transcriptase polymerase chain reaction. Using the patch pipette to both harvest mRNA from an individual neuron and measure neuronal electrical activity, this approach has successfully demonstrated how genes determine function...
Advances in DNA sequencing enable the analysis of the genomes and transcriptomes of single cells and will soon enable single-cell epigenomic and proteomic analyses. Single-cell genomic analysis can reveal genomic variability among individual cells, which can be used to reconstruct cellular ancestries ...
Direct lineage reprogramming involves the conversion of cellular identity. Single-cell technologies are useful for deconstructing the considerable heterogeneity that emerges during lineage conversion. However, lineage relationships are typically lost dur
Single-cell analysis sidesteps this problem by examining gene expression in only one cell. Single-cell methods for both RT-qPCR and RNA-seq have been described. These methods work in one of two ways. Single cells can be isolated with a micromanipulator or into droplets (Imdahl et al., ...
cell requires whole-genome amplification (WGA) by three major methods including degenerate oligonucleotide primed PCR (DOP-PCR), multiple displacement amplification (MDA) and multiple annealing and looping based amplification cycles (MALBAC) [104]. DOP-PCR often yields low genome coverage, which is ...
spin-column format; Small elution volume of 8 µL ready for direct utilization in qRT-PCR reactions; Isolate total RNA including microRNA in a concentrated ready to use format; Isolate high quality total RNA from a variety of sources; RNA can be isolated from a single cell to 100,000 ...
We describe here a direct ex vivo, single-cell-based strategy for the clonotypic analysis of TCRαβ repertoires that utilizes multiplexed panels of TCRα and TCRβ-specific primers in a nested PCR to amplify expressed transcripts from individual, epitope-specific T cells. This strategy yields ...
Next, we transduced Cas9-expressing activated OT-I CD8+T cells (specific for ovalbumin (OVA)) with the scCRISPR library, followed by adoptive transfer to B16-OVA melanoma tumour-bearing mice16. Single-cell sgRNA and transcriptome libraries from donor-derived tumour-infiltrating lymphocytes (TILs...
With CellTag-multi, we present an approach that enables direct capture of heritable random barcodes expressed as polyadenylated transcripts, in both single-cell RNA sequencing and single-cell Assay for Transposase Accessible Chromatin using sequencing assays, allowing for independent clonal tracking of ...