HIGH-THROUGHPUT AUTOMATED STRAIN LIBRARY GENERATOR
A method for automated, high throughput cellular library generation is disclosed. The method includes providing a suspension including transformed cells and plating the transformed cells onto solid surfaces of each of at least one reservoir of a reservoir plate. The solid surfaces can include a liquid growth medium. The reservoir plate is incubated, and after cellular growth has occurred on at least one plated surface of the reservoir plate, a series of automatic steps are performed. The automatically-performed steps include adding disaggregation solution to the reservoir plate, applying a mechanical force, such as a rotational force, to the reservoir plate to produce resuspended cells, and/or collecting the resuspended cells.
1 . A method, comprising:
providing a suspension including a population of cells;
plating cells from the population of cells onto solid surfaces of each of at least one reservoir of a reservoir plate;
incubating the reservoir plate;
after cellular growth has occurred on at least one plated surface of the reservoir plate, automatically:
adding disaggregation solution to the reservoir plate,
applying a mechanical force to the reservoir plate to produce resuspended cells, and
collecting the resuspended cells.
2 . The method of claim 1 , wherein the population of cells includes at least one of transformed cells, transfected cells, culturable bacteria, edited microbes, or natural microbes.
3 . The method of claim 2 , wherein the population of cells includes the transformed cells, the method further comprising introducing DNA to cells to produce the transformed cells, prior to the plating.
4 . The method of claim 1 , further comprising at least one of:
adding beads to the reservoir plate prior to applying the mechanical force to the reservoir plate; or
controlling a temperature of the reservoir plate during the application of the mechanical force, such that the temperature of the reservoir plate is between 4° C. and 40° C.
5 . The method of claim 1 , further comprising at least one of:
automatically transferring the collected resuspended cells to a multi-well plate after the cellular growth has occurred on at least one plated surface of the reservoir plate; or
performing an automated library preparation using the collected resuspended cells.
6 . The method of claim 1 , wherein the disaggregation solution includes at least one of: Triton Z100, Tween-80, Urea, Pluronic™ F-68, Accumax, Accutase, Lipase, a detergent, a protease, an amilase, a cellulose, a glycerol, a microbial growth medium, an antibiotic, a viscous liquid, a positive selection agent or a negative selection agent.
7 . The method of claim 6 , wherein the disaggregation solution includes the microbial growth medium, the glycerol, and the antibiotic.
8 . The method of claim 1 , wherein the mechanical force is at least one of:
a rotational force between 200 RPM and 3,000 RPM; or
applied for a duration of between 10 seconds and 10 minutes.
9 . The method of claim 1 , wherein the solid surfaces of the reservoir plate comprise a liquid growth medium.
10 . The method of claim 1 , wherein the reservoir plate is a prepared reservoir plate, the method further comprising preparing the reservoir plate, to produce the prepared reservoir plate and prior to the plating, by applying a predetermined volume of a growth medium into each well of the reservoir plate.
11 . A method, comprising:
providing a suspension including a population of cells;
plating cells from the population of cells onto solid surfaces of each of at least one reservoir of a reservoir plate;
incubating the reservoir plate;
after spores have grown on at least one plated surface of the reservoir plate, automatically:
adding disaggregation solution to the reservoir plate,
applying a mechanical force to the reservoir plate to produce a spore suspension including at least a portion of the spores, and
collecting the spore suspension.
12 . The method of claim 11 , wherein the solid surfaces of the at least one reservoir of the reservoir plate include an antibiotic.
13 . The method of claim 11 , further comprising performing an automated library preparation based on the spore suspension.
14 . The method of claim 11 , wherein the disaggregation solution includes at least one of: Triton Z100, Tween-80, Urea, Pluronic™ F-68, Accumax, Accutase, Lipase, a detergent, a protease, an amilase, a cellulose, or a glycerol.
15 . The method of claim 11 , wherein the disaggregation solution includes a viscous liquid.
16 . The method of claim 11 , wherein the disaggregation solution includes at least one of a positive selection agent or a negative selection agent.
17 . The method of claim 11 , wherein the disaggregation solution includes a microbial growth medium, glycerol, and an antibiotic.
18 . The method of claim 11 , further comprising adding beads to the reservoir plate prior to applying the mechanical force to the reservoir plate.
19 . The method of claim 11 , wherein the mechanical force is at least one of:
a rotational force between 200 RPM and 3,000 RPM; or
applied for a duration of between 10 seconds and 10 minutes.
20 . The method of claim 11 , further comprising controlling a temperature of the reservoir plate during the application of the mechanical force, such that the temperature of the reservoir plate is between 4° C. and 40° C.