obi-ontology / obi

The Ontology for Biomedical Investigations
http://obi-ontology.org
Creative Commons Attribution 4.0 International
74 stars 24 forks source link

NTR: Clone-seq #877

Open jahilton opened 6 years ago

jahilton commented 6 years ago

editor preferred term: Clone-seq textual definition: A massively-parallel site-directed mutagenesis approach leveraging next-generation sequencing to rapidly and cost-effectively generate a large number of mutant alleles. One colony of each mutagenesis attempt is put into one pool and combine multiple pools through multiplexing for one Illumina sequencing run. definition source for the textual definition: http://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1004819 logical definition: sequencing assay --> Clone-seq

DanBerrios commented 6 years ago

I'm wondering if this is more of an experimental design, rather than a truly novel subtype of sequencing assay. The goal of "Clone-Seq" is "to rapidly and cost-effectively generate a large number of mutant alleles", rather than simply to assay a particular single sample for some feature of that sample. The equipment used to perform the sequencing aspects of "Clone-seq" is standard Illumina equipment. The novel portions are combining site-directed mutagenesis techniques and protein-protein interactome information to generate mutants, then organizing those mutants across standard high-throughput sequencing runs.

DanBerrios commented 5 years ago

@jahilton Do you agree?

bpeters42 commented 5 years ago

I wasn't asked, but I agree :) Also, the definition could be a bit more specific: Is this specific for bacterial cloning ('colony' makes me think that)? Is there something specific about the colony picking? That might make it a specific kind of assay...

On Mon, Oct 15, 2018 at 9:39 AM Dan Berrios notifications@github.com wrote:

@jahilton https://github.com/jahilton Do you agree?

— You are receiving this because you are subscribed to this thread. Reply to this email directly, view it on GitHub https://github.com/obi-ontology/obi/issues/877#issuecomment-429926011, or mute the thread https://github.com/notifications/unsubscribe-auth/ANN9Ijzz6GD1ya83Pf6gw8ljHKtBn5jWks5ulLpIgaJpZM4QVeAA .

-- Bjoern Peters Professor La Jolla Institute for Allergy and Immunology 9420 Athena Circle La Jolla, CA 92037, USA Tel: 858/752-6914 Fax: 858/752-6987 http://www.liai.org/pages/faculty-peters

jahilton commented 5 years ago

Yes, @DanBerrios, I agree with your characterization of the method. And yes, @bpeters42, site-directed mutagenesis, in general, uses a host cell (most often E. coli, I believe) as the vector. The distinction with Clone-seq, specifically, is the pooling/multiplexing aspect