Ontology highlight
ABSTRACT:
SUBMITTER: Tang F
PROVIDER: S-EPMC3847604 | biostudies-literature | 2010 Mar
REPOSITORIES: biostudies-literature
Nature protocols 20100225 3
We describe here a protocol for digital transcriptome analysis in a single mouse oocyte and blastomere using a deep-sequencing approach. In this method, individual cells are isolated and transferred into lysate buffer by mouth pipette, followed by reverse transcription carried out directly on the whole cell lysate. Free primers are removed by exonuclease I and a poly(A) tail is added to the 3' end of the first-strand cDNAs by terminal deoxynucleotidyl transferase. Single-cell cDNAs are then ampl ...[more]