Patent US4746842A successfully refreshed from Google Patents.
US5124444A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Lactam-containing compositions and methods useful for the extraction of nucleic acids
Assignee: Becton Dickinson and Co
•
Filing: 1989-07-24
•
Granted: 1992-06-23
•
Expires: 2009-07-24
Abstract
Abstract This invention relates to novel methods for the extraction of nucleic acid. In particular methods are described for isolating nucleic acid from a sample containing a complex biological mixture of nucleic acid and non-nucleic acids wherein the sample is combined with an extraction solution comprising a lactam and then the nucleic acid material is isolated from the resulting combined solution. The resulting combined solution is mixed and becomes biphasic and the nucleic acid material is isolated from the aqueous phase by precipitation with ethanol. The lactam is preferably about 5 to about 70% of the extraction solution and is most preferably 2-pyrrolidone, N-ethyl-2-pyrrolidone, N-cyclohexyl-2-pyrrolidone, N-dodecyl-2-pyrrolidone, N-methyl-2-pyrrolidone, N-hydroxyethyl-2-pyrrolidone, N-methyl-2-piperidone, 2-ε-caprolactam, N-methyl-2-caprolactam, 2-piperidone or N-(4-hydroxybenzyl)pyrrolidone. Methods for selectively isolating DNA, ribosomal RNA and plasmid DNA are also disclosed.
Inventors
- 👤 Jeffrey Van Ness
- 👤 Nicolaas Vermuelen
- 👤 B. Melina Cimler
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 21:36
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6832; C12Q1/6806; Y; Y10; Y10T; Y10T436/00; Y10T436/25; Y10T436/25375; Y10T436/255
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
75 words
A method for isolating nucleic acid from a sample containing a complex biological mixture of nucleic acids and non-nucleic acids, comprising: combining the sample with an extraction solution comprising at least one monomeric lactam at a concentration of about 5% to about 70% of the total volume of the extraction solution, wherein the lactam is capable of forming a biphasic solution after combination with the sample; and isolating nucleic acid from the resulting combined solution.
Claim 2
Independent Claim
75 words
A method for isolating nucleic acid from a sample containing a complex biological mixture of nucleic acids and non-nucleic acids, comprising: combining the sample with an extraction solution comprising at least one monomeric lactam at a concentration of about 5% to about 70% of the total volume of the extraction solution, wherein the lactam is capable of forming a biphasic solution after combination with the sample; and isolating nucleic acid from the resulting combined solution.
Claim 3
Dependent Claim
170 words
The method of claim 1, wherein the lactam is selected from the group of compounds having the formula: ##STR5## wherein n=3 to 8 (with the total number of R substituents being 3 to 8), and R 1 is selected from the group consisting of: hydrogen, alkyl of 1 to 20 carbons, aryl of 6 to 10 carbons, arylalkyl of 7 to 20 carbons, alkylcarboxyamide of 1 to 20 carbons, hydroxyarylalkyl of 7 to 20 carbons, N-mono-substituted alkylcarboxyamide, N,N-dialkyl-substituted alkylcarboxyamide, alkylcarboxylate of 2 to 20 carbons, acyl of 2 to 20 carbons, cycloalkane of 4 to 10 carbons, hydroxyalkyl, and cyanoalkyl; and where the R substituents can be the same or different and selected from a group consisting of hydrogen, halogen and alkyl of 1 to 5 carbons such that the total carbon number for all R substituents does not exceed 20; with the further provision that the total carbon number of the R 1 substituent does not exceed 20 and where at least one R designated substituent must be hydrogen.
Claim 4
Dependent Claim
9 words
The method of claim 2, wherein n=3 to 6.
Claim 5
Dependent Claim
13 words
The method of claim 2, wherein at least one R substituent is methyl.
Claim 6
Dependent Claim
18 words
The method of claim 2, wherein the R 1 substituent and all of the R substituents are hydrogen.
Claim 7
Dependent Claim
22 words
The method of claim 2, wherein the R 1 substituent is other than hydrogen and all of the R substituents are hydrogen.
Claim 8
Dependent Claim
18 words
The method of claim 2, wherein the R 1 substituent is a cycloalkane of 4 to 7 carbons.
Claim 9
Dependent Claim
16 words
The method of claim 2, wherein the R 1 substituent is --CH 2 --CH 2 OH.
Claim 10
Dependent Claim
12 words
The method of claim 2, wherein the R 1 substituent is hydroxyarylalkyl.
Claim 11
Dependent Claim
12 words
The method of claim 2, wherein the R 1 substituent is dodecyl.
Claim 12
Dependent Claim
12 words
The method of claim 2, wherein the R 1 substituent is methyl.
Claim 13
Dependent Claim
29 words
The method of claim 1, wherein the lactam is a compound selected from the group consisting of 2-pyrrolidone, N-ethyl-2-pyrrolidone, N-cyclohexyl-2pyrrolidone, N-dodecyl-2-pyrrolidone, N-methyl-2-pyrrolidone, N-hydroxyethyl-2-pyrrolidone, N-methyl-2-piperidone, 2-ε-caprolactam, N-methyl-2-caprolactam, 2-piperidone and N-(4-hydroxybenzyl)pyrrolidone.
Claim 14
Dependent Claim
12 words
The method of claim 1, wherein the nucleic acid is plasmid DNA.
Claim 15
Dependent Claim
25 words
The method of claim 1, wherein the resulting combined solution is further mixed and allowed to become biphasic before the nucleic acid material is isolated.
Claim 16
Dependent Claim
20 words
The method of claim 14, wherein the nucleic acid material is isolated by precipitation with ethanol from an aqueous phase.
Claim 17
Dependent Claim
15 words
The method of claim 1, wherein the resulting combined solution further comprises a chaotropic agent.
Claim 18
Dependent Claim
16 words
The method of claim 1, wherein the sample is also combined with a solution comprising GnHCl.
Claim 19
Dependent Claim
35 words
A composition for the extraction of nucleic acid comprising: a buffer solution; and at least one monomeric lactam at a concentration of about 5% to about 70% of the total volume of the buffer solution.
Claim 20
Dependent Claim
35 words
A composition for the extraction of nucleic acid comprising: a buffer solution; and at least one monomeric lactam at a concentration of about 5% to about 70% of the total volume of the buffer solution.
Claim 21
Dependent Claim
37 words
A nucleic acid extraction kit comprising, in one or more containers: a buffer solution; and at least one monomeric lactam at a concentration of about 5% to about 70% of the total volume of the buffer solution.
Claim 22
Dependent Claim
37 words
A nucleic acid extraction kit comprising, in one or more containers: a buffer solution; and at least one monomeric lactam at a concentration of about 5% to about 70% of the total volume of the buffer solution.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-06-23 | 1995-12-23 | 1996-06-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-06-23 | 1999-12-23 | 2000-06-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-06-23 | 2003-12-23 | 2004-06-23 | $7700.00 | Window Closed / Lapsed |