BEGIN:VCALENDAR VERSION:2.0 PRODID:-//Talks.cam//talks.cam.ac.uk// X-WR-CALNAME:Talks.cam BEGIN:VEVENT SUMMARY:Single Cell Seminar: "\;Using single cell transcriptomic signa tures to understand tumour bulk RNA-seq"\; &\; "\;Detecting ger mline mutations in single gametes by preserving molecular identity during amplification"\; - Matthew Young (Behjati group) &\; Chris Laumer (Marioni/Birney groups) DTSTART:20181019T140000Z DTEND:20181019T150000Z UID:TALK106315@talks.cam.ac.uk CONTACT:Lia Chappell DESCRIPTION:Please come along the monthly campus Single Cell Seminar serie s. All are welcome\, including colleagues from Cambridge (though email me so we can book them in as a visitor with security). This month we have two exciting talks\, please join us C302 from 3.00 - 4.00pm THIS FRIDAY (19th October)\n\n1) Matthew Young (Behjati group): "Using single cell transcri ptomic signatures to understand tumour bulk RNA-seq"\n\n2) Chris Laumer (M arioni/Birney groups): "Detecting germline mutations in single gametes by preserving molecular identity during amplification”\n\n\nChris’s abstr act: \nGeneticists study the generation and transmission of new genetic va riation by comparing deeply resequenced genomes of parents and offspring\; variants found in offspring but neither parent are mutations. However\, t his pedigree sequencing design is limited by family size\, and might mask variation in the germline mutation process within and between individuals. Single cell genomics could provide an alternative approach\, allowing mut ations to be discovered in large samples of gametes from a single individu al. However\, this field is limited by the enzymatic pre-amplification pro cess typically used\, which breaks the link between sequencing read and mo lecule of origin\, allowing false positive mutations and undetectable alle lic dropouts to predominate. Here\, we present efforts to overcome these c hallenges through a novel protocol that capitalizes on the concept of uniq ue molecular identity. The method uses restriction enzymes to fragment the naked genome of a lysed single cell\, ligates adapters with double-strand ed molecular barcodes to each fragment\, and then linearly amplifies the D NA from both strands independently\, all in succession without intermediat e clean-up steps. Sequencing reads generated from such libraries are by de sign unambiguously assignable to their original molecule of origin\, there fore permitting sequencing and amplification errors to be corrected by for ming a computational consensus of duplicate reads\, and also allowing coun ting of molecules captured\, facilitating detection of allelic and strand dropouts. It can also be modified to produce reduced representation librar ies\, permitting the pooling of many cells on a single sequencing lane. We are applying this method to sequence single gametes from a hermaphroditic marine invertebrate model\, Ciona intestinalis\, which has a small genome (~160 Mbp)\, and from which gametes and embryos can be easily obtained an d manipulated. Our data will test the hypothesis that individual gametes c an serve as reasonable proxies for successfully developed offspring in stu dying germline mutations. LOCATION:C302\, Sulston Building\, Wellcome Genome Campus\, Hinxton. CB10 1SA END:VEVENT END:VCALENDAR