Method of purifying nucleic acids and kit for purifying nucleic acids

Patent metadata

Field	Value
Publication number	US-10023860-B2
Application number	US-201615295720-A
Country	US
Kind code	B2
Filing date	Oct 17, 2016
Priority date	Sep 13, 2011
Publication date	Jul 17, 2018
Grant date	Jul 17, 2018

Abstract

[Object] To provide a method of purifying nucleic acids where the operation is simple and the nucleic acids can be extracted in a short time with high efficiency.
[Solving Means] A method of purifying nucleic acids including the step of adsorbing substances in a sample containing nucleic acids with an ion exchange resin 10 including a positive ion exchange resin and a negative ion exchange resin. As the positive ion exchange resin, a first positive ion exchange resin and a second positive ion exchange resin having an exclusion limit molecular weight lower than that of the first positive ion exchange resin may be used.

First claim

The invention claimed is:

1. A method of amplifying nucleic acids, the method comprising:
adsorbing substances in a sample containing nucleic acids with an ion exchange resin including a positive ion exchange resin and a negative ion exchange resin thereby purifying the nucleic acids, wherein the positive ion exchange resin includes a first positive ion exchange resin and a second positive ion exchange resin having an exclusion limit molecular weight lower than that of the first positive ion exchange resin, and
amplifying nucleic acids by adding a nucleic acid amplification reagent to the purified nucleic acids.

2. The method of claim 1 , wherein the substances are adsorbed by the first positive ion exchange resin and then are further adsorbed by the negative ion exchange resin and the second positive ion exchange resin.

3. The method of claim 2 , wherein the sample is flowed into a column including the first positive ion exchange resin in an upper layer and the negative ion exchange resin and the second positive ion exchange resin in a lower layer from an upper layer side.

4. The method of claim 3 , wherein the step is for adsorbing the substances included in the sample that is diluted with a buffer solution by the ion exchange resin, and the buffer solution has a pH of 4.0 to 8.0.

5. The method of claim 4 , wherein the positive ion exchange resin is a strong acidic positive ion exchange resin.

6. The method of claim 5 , wherein a counter ion of the first positive ion exchange resin is Na + .

7. The method of claim 6 , wherein a counter ion of the second positive ion exchange resin is H + .

8. The method of claim 7 , wherein the negative ion exchange resin is a strong basic negative ion exchange resin.

9. The method of claim 8 , wherein a counter ion of the negative ion exchange resin is OH − .

10. The method of claim 9 , wherein a percentage of an ion exchange capacity of the negative ion exchange resin to the second positive ion exchange resin is 50% to 150%.

11. The method of claim 10 , wherein a nonionic surfactant and/or a nonionic hydrophilic polymer compound is used to adsorb the substances.

12. The method of claim 11 , wherein the substances are foreign substances containing at least a protein and a metal salt.

13. The method of claim 1 , further comprising the step of ultrasonically treating the sample including nucleic acids.

14. The method of claim 1 , further comprising the step of concentrating the nucleic acids by blocking the nucleic acids migrated by electrophoresis.

15. The method of claim 14 , wherein the electrophoresis is carried out on the nucleic acids into which an intercalator having an anionic functional group is inserted.

16. The method of claim 15 , wherein the electrophoresis is carried out on the nucleic acids by mixing the sample, a compound having a functional group that is reacted with a carboxyl group of the substances included in the sample by dehydration condensation, and a condensation agent of the dehydration condensation reaction.

17. A kit for amplifying nucleic acids, the kit comprising:
a positive ion exchange resin;
a negative ion exchange resin;
a nucleic acid amplification reagent; and
a nucleic acid purifying instrument internally holding the positive ion exchange resin and the negative ion exchange resin for distributing a sample including nucleic acids, wherein the positive ion exchange resin includes a first positive ion exchange resin and a second positive ion exchange resin having an exclusion limit molecular weight lower than that of the first positive ion exchange resin.

18. The kit of claim 17 , wherein the first positive ion exchange resin is a strong acidic positive ion exchange resin and the second positive ion exchange resin is a strong acidic positive ion exchange resin.

19. The kit of claim 18 , wherein a counter ion of the first positive ion exchange resin is Na + .

20. The kit of claim 18 , wherein a counter ion of the second positive ion exchange resin is H + .

21. The kit of claim 17 , wherein a percentage of an ion exchange capacity of the negative ion exchange resin to the second positive ion exchange resin is 50% to 150%.

Assignees

• Sony Corp

Inventors

• Nakamura Tomohiko
• Sakamoto Naohisa
• Yotoriyama Tasuku
• Ito Kazumine

Classifications

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• B01D15/363

  Anion-exchange · CPC title

• B01D15/362

  Cation-exchange · CPC title

• B01D15/34

  Size-selective separation, e.g. size-exclusion chromatography; Gel filtration; Permeation · CPC title

• G01N2030/8827

  involving nucleic acids · CPC title