公开(公告)号：US11732257B2

公开(公告)日：2023-08-22

申请号：US16758485

申请日：2018-10-23

申请人： Massachusetts Institute of Technology ,  The General Hospital Corporation

发明人： Peter van Galen ,  Volker Hovestadt ,  Travis Hughes ,  Marc H. Wadsworth, II ,  Bradley Bernstein ,  Alexander K. Shalek ,  Todd M. Gierahn ,  J. Christopher Love ,  Ang A. Tu

IPC分类号： C12P19/34 ,  C12N15/10 ,  C12Q1/686 ,  C12Q1/6886 ,  C12Q1/6888

CPC分类号： C12N15/1065 ,  C12Q1/686 ,  C12Q1/6886 ,  C12Q1/6888 ,  C12Q2600/158

摘要： The present invention relates to methods of detecting region(s) of interest in a gene comprising a polyA tail. The region(s) of interest can include gene(s), region(s), mutation(s), deletion(s), insertion(s), indel(s), and/or translocation(s). The region(s) can be greater than or less than 1 kilobases from the polyA tail. Methods can include forming a library of single cell transcripts comprising the region(s) in close proximity to a cell barcode and a unique molecular identifier (UMI). Methods for distinguishing cells by genotype can include amplifying the transcripts using PCR methods and detecting the cell barcode and UMI using single cell sequencing methods. Transcripts can be enriched using tagged region-specific PCR primers. Cell barcodes can be brought into close proximity to the region(s) by circularizing the transcripts. Sequencing of the transcripts can include using primer binding sites added during PCR amplification and library indexes for multiplexed sequencing.

公开(公告)号：US12071663B2

公开(公告)日：2024-08-27

申请号：US16070505

申请日：2017-01-17

申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY

发明人： Todd M. Gierahn ,  J. Christopher Love ,  Travis K. Hughes ,  Marc H. Wadsworth, II ,  Alexander K. Shalek ,  Shaina Carroll

IPC分类号： B01J19/00 ,  C12Q1/6874

CPC分类号： C12Q1/6874 ,  B01J19/0046 ,  B01J2219/00572 ,  C12Q1/6874 ,  C12Q2537/143 ,  C12Q2563/149 ,  C12Q2563/179 ,  C12Q2565/514

摘要： The present application provides a method of assembling a container for one or multiple parallel steps of biochemical analysis on one or more cells comprising performing molecular bonding of a porous membrane on an apical or basal surface of an array having a plurality of wells, wherein the molecular bonding substantially isolates each well from adjacent wells.