Method for capturing a nucleic acid

• US 5,939,261 A
• Filed: 06/24/1997
• Issued: 08/17/1999
• Est. Priority Date: 06/24/1997
• Status: Expired due to Fees

• Current Assignee: Sarnoff Corporation (SRI International, Inc.)
• Original Assignee: Sarnoff Corporation (SRI International, Inc.)
• Inventors: Kumar, Rajan, Loewy, Zvi
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US08/881,282
• Time in Patent Office: 784 Days
• Field of Search: 435/5, 435/6, 536/23.1, 536/24.3, 935/76, 935/77, 935/78
• US Class Current: 435/6.11
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase

Abstract

The present invention relates to a method for capturing a class of nucleic acids comprising (a) providing a population of nucleic acids, wherein the population comprises the class; (b) binding a probe moiety to nucleic acids of the class, thereby forming one or more complexes, wherein the probe moiety is attached to a substrate; and (c) capturing the complex. The method can be implemented in certain embodiments in a structure comprised of one or more reservoirs, preferably wherein the reservoirs include one or more second chambers that are in communication with a first chamber. Preferably, the present invention is used to capture the class of nucleic acids that comprise regulatory elements included in a population of nucleic acids isolated from a cell, tissue, or organism. An alternative preferred embodiment of the present invention involves the capture of the class of nucleic acids that comprises certain repetitive elements included in a population of nucleic acids derived from an organism.

Claims

1. A method for capturing a class of nucleic acids in a device, the method comprising:

a. furnishing a device for relaying liquids in channels comprising the channels and one or more chambers;

b. combining in a first chamber of the device (1) a population of nucleic acids comprised of nucleic acids of the class and (2) a probe that binds to the class of nucleic acids, the probe having an attachment moiety suitable for attaching to a first substrate on at least one paramagnetic or superparamagnetic microparticle, wherein the probe is a DNA binding protein or a repetitive nucleic acid element;

c. forming one or more complexes of the probe and the class of nucleic acids;

d. capturing the class of nucleic acids from the population of nucleic acids in the device by capturing the complex bound to the first substrate via an attachment of the moiety to the first substrate; and

e. separating the class of nucleic acids bound to the first substrate from the unbound nucleic acids by (1) magnetically retaining the microparticles in one said chamber and moving through at least one of said channels liquid containing the unbound nucleic acids to another said chamber or (2) magnetically moving through at least one of said channels the microparticles from one said chamber to another.

2. The method of claim 1, wherein the nucleic acids of the population are RNA or DNA.

3. The method of claim 2, wherein the nucleic acids of the population are nuclease products or amplicons.

4. The method of claim 1, wherein the first chamber has a volume of from about 0.05 μ

l to about 250 μ

l.

5. The method of claim 1, wherein the microparticle is superparamagnetic.

6. The method of claim 1, wherein the probe binds preferentially to a specific sequence.

7. The method of claim 1, wherein the probe is a DNA binding protein which is Sp1, C-FOS, WT-1, or CRO.

8. The method of claim 1, wherein the nucleic acid, the probe, or first substrate includes an indicator moiety, provided that distinguishable indicator moieties are used when at least two of the nucleic acid, the probe and first substrate include an indicator moiety.

9. The method of claim 1, further comprising detecting the complex, wherein the detecting of the complex comprises exposing a detector to the complex, wherein the detector is a detector of radiation, fluorescence, or light.

10. The method of claim 1, wherein the probe is distributed to two or more first chambers, wherein the probe in one first chamber binds to a different class of nucleic acids than does the probe included in a different first chamber.

11. The method of claim 10, wherein the probe in each of the first chambers is placed in contact with the population of nucleic acids.

12. The method of claim 1, wherein the DNA binding protein preferentially binds to regulatory elements included in the population of nucleic acids.

13. The method of claim 1, further comprising:

f. forming a second complex between (1) a second probe that binds to a second class of nucleic acids, the second probe having a second attachment moiety, and (2) the second class of nucleic acids; and

g. capturing the second complex bound to a second substrate via the second attachment moiety.

14. The method of claim 13, wherein the second probe or the second substrate includes an indicator moiety, provided that distinguishable indicator moieties are used when both the second probe and the second substrate include an indicator moiety.

15. The method of claim 13, wherein aliquots of the second substrate-attached nucleic acids of the class are distributed to two or more first chambers.

16. The method of claim 15, wherein the aliquots in the respective first chambers are placed in contact with the probe, further wherein the probe in each of the chambers binds to different classes of nucleic acid.

17. The method of claim 1, further comprising.

e'"'"'. magnetically moving the microparticle with the captured complex to a second chamber which is in fluid communication with the first chamber within the device to separate the class of nucleic acids from the remainder of the population of nucleic acids.

18. The method of claim 1, wherein the first substrate has an attached binding moiety, wherein the binding moiety and the attachment moiety preferentially or exclusively bind to each other to form a binding pair, thereby forming the attachment.

19. The method of claim 18, further comprising different substrates each having an appropriate binding moiety that attaches to a different probe, each probe having an appropriate corresponding attachment moiety.