Abstract: Methods to emulsify cells and/or mRNA with reverse transcriptase at a temperature such that the reverse transcriptase begins making cDNA during the emulsification

Abstract: This disclosure provides methods and systems for single-cell analysis, including single-cell transcriptome analysis, of target cells without microfluidic devices. The disclosed methods involve the use of template particles to template the formation of monodisperse droplets to generally capture a single target cell from a population of cells in an encapsulation, derive a plurality of distinct mRNA molecules from the single target cell, and quantify the distinct mRNA molecules to generate an expression profile.

Abstract: The present disclosure provides a method of hairpin primer design and targeted amplification thereof for creation of targeted sequencing libraries. Generally, the present methods allow for simultaneous construction of targeted sequencing libraries for multiple targeted nucleic acid sequences within a single sample. The presently disclosed methods generate efficient and specific target sequence amplification while avoiding, or significantly reducing, non-specific interaction of multiplex primers, non-specific amplification of sequences due to random priming from molecular “tags” (such as Molecular Identifiers (“MI”) barcodes), and unintentional interactions between gene-specific and universal primers.