Automated method for isolation, selection and/or detection of microorganisms or cells comprised in a solution

Patent metadata

Field	Value
Publication number	US-11225682-B2
Application number	US-201615767481-A
Country	US
Kind code	B2
Filing date	Oct 12, 2016
Priority date	Oct 12, 2015
Publication date	Jan 18, 2022
Grant date	Jan 18, 2022

Abstract

The present invention is concerned with an automated method for obtaining at least one discrete colony from microorganisms or cells comprised in a solution. This method includes a step where acoustic liquid transfer is employed. The present invention further relates to the use of an acoustic liquid transfer device for obtaining at least one discrete colony from micro organisms or cells comprised in a solution. The present invention also relates to an automated method for determining the presence and/or quantity of microorganisms or cells potentially comprised in a solution and to the use of an acoustic liquid transfer device for determining the presence and/or quantity of microorganisms or cells potentially comprised in a solution.

First claim

The invention claimed is:

1. An automated method for obtaining at least one discrete colony from microorganisms or cells comprised in a solution, the method comprising the steps of:
a) transforming or transducing microorganisms or cells with an exogenous nucleic acid comprising selection marker and providing a solution comprising the microorganisms or cells;
b) providing a solid medium suitable for growth of said microorganisms or cells selective for said microorganisms or cells comprising the exogenous nucleic acid;
c) dispensing at least two differently defined volumes of said solution provided in step a) onto said solid medium provided in step b) using acoustic liquid transfer, said at least two differently defined volumes being selected from a range of from 1.0 nl to 100 μl, wherein said acoustic liquid transfer comprises using a transducer which generates and transfers acoustic energy, and wherein said transducer does not directly contact said solution, wherein the at least two differently defined volumes are dispensed at a distance of at least 100 μm apart; and
d) incubating said solid medium under conditions suitable for growth of said microorganisms or cells and thus obtaining at least one discrete colony of living microorganisms or cells comprising the exogenous nucleic acid.

2. The method according to claim 1 , wherein said solution comprises an undefined number of colony-forming units.

3. The method according to claim 1 , wherein in total a volume of said solution comprising microorganisms or cells from a range of from 100 nl to 50 μl is dispensed onto said solid medium.

4. The method according to claim 1 , wherein said at least two differently defined volumes are selected from a lower end and an upper end of said range close to an end of the volume range and opposite from the other volume.

5. The method according to claim 1 , wherein said method is carried out in parallel with at least two solutions comprising microorganisms or cells.

6. The method according to claim 1 , wherein said method is carried out on a culture plate containing a single well.

7. The method of claim 1 , wherein the solution comprises an unknown number of microorganisms or cells.

8. The method of claim 1 , wherein the microorganisms or cells are bacterial cells.

9. The method of claim 8 , wherein the bacterial cells are E. coli.

10. The method of claim 8 , wherein the selection marker provides resistance to an antibiotic.

11. The method of claim 10 , wherein the antibiotic is ampicillin and/or kanamycin.

Assignees

• Curevac Ag

Inventors

• Reichert Isabel
• Eber Fabian Johannes
• Mayer Christian
• Wochner Aniela

Classifications

• C12Q1/20

  using multifield media · CPC title

• C12Q1/24 Primary

  Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title