Modular flow cells and methods of sequencing

• US 10,150,994 B2
• Filed: 07/20/2016
• Issued: 12/11/2018
• Est. Priority Date: 07/22/2015
• Status: Active Grant

• Current Assignee: Qiagen GmbH (Qiagen NV)
• Original Assignee: QIAGEN Instruments AG (Qiagen NV), Qiagen Waltham, Inc.
• Inventors: Olejnik, Jerzy, Zimmermann, Dirk
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US15/215,234
• Publication Number:
  US 20170022555A1
• Time in Patent Office: 874 Days
• Field of Search: 435 61, 435 911, 433 681, 422 681
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2523/107 Chemical cleaving agents
• CPC Class Codes: C12Q 2525/186 incorporating a non-extendable or blocking moiety
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2565/629 being a microfluidic device

Abstract

Modular flow cells, devices with modular flow cells, and methods of sequencing using modular flow cells, as well as systems and kits including modular flow cells, are described, permitting sequencing wherein less than the full capacity for sequencing is desired.

Claims

1. A method of incorporating labeled nucleotides into nucleic acid, comprising:

a) providing

i) a modular flow cell comprising a plurality of modules, wherein said modules are moveable and removable inserts, each module comprising biomolecules bound to a surface of a lane or fluidic channel, said plurality of modules sharing an input and output port, wherein each module comprises a fluidic channel and the fluidic channel of one module is blocked so that no solutions enter said blocked fluidic channel,

ii) a first solution comprising polymerase and a plurality of nucleotide analogues wherein at least one nucleotide analogue is labeled with a unique label and contains a removable chemical moiety capping the 3′

-OH group, and

iii) a second solution comprising a cleaving agent;

b) introducing said first solution into at least one module of said plurality of modules under conditions wherein a first nucleotide analogue is incorporated by said polymerase;

c) detecting the label of the incorporated nucleotide analogue; and

d) introducing said second solution into said at least one module under conditions such that the chemical moiety of the incorporated nucleotide analogue capping the 3′

-OH group is removed by said cleaving agent.

2. The method of claim 1, wherein the fluidic channel of said one module is blocked with a plug.

3. The method of claim 1, wherein said biomolecules comprise primers for sequencing.

4. A method of incorporating labeled nucleotides into nucleic acid, comprising:

a) providing

i) a modular flow cell comprising a plurality of modules, wherein said modules are moveable and removable inserts, each module comprising biomolecules bound to a surface of a fluidic channel, each fluidic channel having a separate input and output port, wherein the fluidic channel of one module is blocked so that no solutions enter said blocked fluidic channel,

ii) a first solution comprising polymerase and a plurality of nucleotide analogues wherein at least one nucleotide analogue is labeled with a unique label and contains a removable chemical moiety capping the 3′

-OH group, and

iii) a second solution comprising a cleaving agent;

b) introducing said first solution into at least one module of said plurality of modules under conditions wherein a first nucleotide analogue is incorporated by said polymerase;

c) detecting the label of the incorporated nucleotide analogue; and

d) introducing said second solution into said at least one module under conditions such that the chemical moiety of the incorporated nucleotide analogue capping the 3′

-OH group is removed by said cleaving agent.

5. The method of claim 4, wherein the fluidic channel of said one module is blocked with a plug.

6. The method of claim 4, wherein said biomolecules comprise primers.

7. A method of incorporating labeled nucleotides into nucleic acid, comprising:

a) providing

i) a modular flow cell comprising four modules, wherein said modules are moveable and removable inserts, at least one of which comprise biomolecules bound to a surface of a fluidic channel, wherein one module comprises no biomolecules in the fluidic channel, and no sequencing takes place in that channel;

ii) a first solution comprising polymerase and a plurality of nucleotide analogues wherein each nucleotide analogue is labeled with a unique label and contains a removable chemical moiety capping the 3′

-OH group, and

iii) a second solution comprising a cleaving agent;

b) introducing said first solution into said four modules under conditions wherein a first nucleotide analogue is incorporated by said polymerase in at least one of said four modules;

c) detecting the label of the incorporated nucleotide analogue; and

d) introducing said second solution into said at least one module under conditions such that the chemical moiety of the incorporated nucleotide analogue capping the 3′

-OH group is removed by said cleaving agent.

8. The method of claim 7, wherein said biomolecules comprise primers for sequencing.