Instruments, modules, and methods for improved detection of edited sequences in live cells

A method for creating edited cell libraries comprising:

• providing cells to be edited;
• providing nucleic acids to edit the cells to be edited;
• providing a growth module, a transformation module and a singulation or substantial singulation, growth, induction of editing, and normalization or cherry-picking (SWIIN) module, wherein the SWIIN module comprises a singulation assembly, wherein the singulation assembly comprises:
• a retentate member comprising an upper surface and a lower surface, wherein the retentate member comprises at least one retentate distribution channel which traverses the retentate member from its upper surface to its lower surface and for 70% to 95% of a length of retentate member;
• wherein the lower surface of the retentate member comprises a first portion of a serpentine channel;
• wherein the retentate member further comprises one or more retentate member ports configured to supply cells and fluid to and remove cells and fluid from the retentate member;
• and wherein the retentate member ports are fluidically-connected to the at least one retentate distribution channel and retentate flow directors;
• the permeate member comprising the upper surface and a lower surface, wherein the permeate member comprises at least one permeate distribution channel which traverses the permeate member from its lower surface to its upper surface and for 70% to 95% of a length of permeate member;
• wherein the upper surface of the permeate member comprises a second portion of the serpentine channel;
• wherein the permeate member further comprises one or more permeate member ports configured to supply fluid to and remove fluid from the permeate member;
• wherein the permeate member ports are fluidically-connected to the at least one permeate distribution channel and permeate flow directors;
• and wherein the first and second portions of the serpentine channel mate to create the serpentine channel;
• a gasket disposed between the retentate member and the permeate member;
• and
• means to couple the retentate member and permeate member;
• transferring the cells to be edited to the growth module;
• growing the cells to be edited;
• transferring the grown cells from the growth module to the transformation module;
• transferring the nucleic acids to the transformation module;
• transforming the grown cells with the nucleic acids;
• singulating the transformed cells in the SWIIN module;
• and
• providing conditions in the SWIIN module to edit the cells, thereby creating an edited cell library.