Dicationic perylene dye for precipitation and quantitation of DNA

• US 5,811,307 A
• Filed: 06/11/1997
• Issued: 09/22/1998
• Est. Priority Date: 01/11/1996
• Status: Expired due to Term

• Current Assignee: Florida State University (State University System of Florida)
• Original Assignee: Florida State University (State University System of Florida)
• Inventors: Liu, Zhi-Ren, Rill, Randolph L.
• Primary Examiner(s): ALEXANDER, LYLE
• Application Number:
  US08/873,222
• Time in Patent Office: 468 Days
• Field of Search: 436/86-93, 436/175, 436/63, 436/71, 436/94
• US Class Current: 436/94
• CPC Class Codes: C09B 5/62 Cyclic imides or amidines of peri-dicarboxylic acids of the anthracene, benzanthrene, or perylene series
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials

Abstract

The novel dicationic dye N,N'"'"'-bis 3,3'"'"'-(dimethylamino)propylamine!-3,4,9,10-perylenetetracarboxylic diimide (DAPER) is prepared by reacting 3,4,9,10-perylenetetracarboxylic dianhydride with N,N-dimethylaminopropylamine. The dye DAPER is particularly useful in a rapid, sensitive method for the precipitation, recovery and quantitation of DNA from solutions containing DNA, which method involves the steps of (a) adding the dye DAPER to a solution containing DNA and allowing the resulting mixture to equilibrate; (b) collecting the resulting precipitate; (c) removing weakly bound DAPER from the precipitate; (d) extracting DAPER therefrom by the addition of a solvent therefor; and (e) determining the amount of DNA precipitated by measuring either the absorbance or fluorescence of DAPER in the solvent extract from step (d).

Claims

1. A process for the precipitation, recovery and quantitation of DNA from solutions containing DNA which comprises the steps of:

a. adding N,N'"'"'-bis- 3,4,9,10-perylenetetracarboxylic diimide to a solution containing DNA and allowing the resulting mixture to equilibrate;

b. collecting the resulting precipitate;

c. removing weakly bound excess N,N'"'"'-bis-3,4,9,10-perylenetetracarboxylic diimide from said precipitate;

d. extracting N,N'"'"'-bis-3,4,9,10-perylenetetracarboxylic diimide from said precipitate by the addition of a solvent for N,N'"'"'-bis-3,4,9,10-perylenetetracarboxylic diimide; and

e. determining the amount of DNA precipitated by measuring either the absorbance or fluorescence of N,N'"'"'-bis-3,4,9,10-perylenetetracarboxylic diimide in the solvent extract from step d.

2. A process as set forth in claim 1 wherein said solvent is selected from the group consisting of chloroform, phenol:

chloroform and dimethylformamide.

3. A process as set forth in claim 1 wherein N,N'"'"'-bis-3,4,9,10-perylenetetracarboxylic diimide is removed from said precipitate in step c by washing the precipitate with water.