Bio-rad Laboratories, Inc.

Xiangdong Meng

Man Cheng

Steven Okino

Yanhong Kong

Xiaosong Gong

Cindy C T Wan

Jan Zur Megede

Steven T Okino

The subject application pertains to compositions and methods for enhancing reverse transcriptase (RT) activity and/or reducing the inhibition of RT by inhibitors, such as formalin, tannic acid and/or heparin. In some embodiments, RT inhibition is reduced by the addition of potassium glutamate, histidine hydrochloride monohydrate, poloxamer 188, or any combination thereof to a reaction mixture comprising a polymerase. In other embodiments, RT is enhanced through the addition of a polyvinyl sulfonic acid sodium salt (PVSA) to a reaction mixture. The subject application also provides oligonucleotide primers for use in the reverse transcription of target sequences and its enhancement. These primers have high GC content or low GC content. Methods of using a RT inhibition reducer or a RT enhancer in a composition with an RNA template and RT improves RT yield, RT sensitivity, or RT tolerance to various chemicals are also provided.

Compositions and methods for enhancing reverse transcriptase activity and/or reducing the inhibition of reverse transcriptase

Mutant Type-A DNA polymerases having increased resistance to one or more polymerization activity inhibitors are provided.

Mutant DNA polymerases

Methods of determining polymerase fidelity are provided. In one embodiment, the method comprises filling a gapped plasmid with a polymerase to form a gap-filled plasmid, wherein the gap-filled plasmid comprises a gene encoding an protein that is functional or non-functional depending on the polymerase fidelity; forming a plurality of partitions from a solution comprising the gap-filled plasmid and a label for detecting the presence of the plasmid; detecting the presence of the gap-filled plasmid in one or more of the partitions; and determining the fidelity of the polymerase by determining a ratio of partitions containing the gene encoding a functional protein to partitions containing a gene encoding a non-functional protein.

Digital polymerase fidelity assay

Filtering small nucleic acids using permeabilized cells and methods for using the filtering to detect genomic DNA accessibility are described.

Growing community of inventors

Albany, CA, United States of America

Xiangdong Meng