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Compositions and methods for the release of nucleic acid molecules from solid matrices

a nucleic acid and solid matrices technology, applied in biochemistry apparatus and processes, organic chemistry, sugar derivatives, etc., can solve the problems of hiv, a number of infectious agents, and the loss of viability of agricultural products prepared from genetically modified plants, so as to facilitate the release of nucleic acid molecules

Inactive Publication Date: 2005-01-13
INVITROGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0015] The present invention relates to compositions and methods for the removal of nucleic acid molecules (e.g., DNA) from solid matrices. In particular, the meth...

Problems solved by technology

Further, a number of consumer groups, as well as governmental organizations, have objected to the sale of agricultural products prepared from genetically modified plants.
More specifically, HIV, as well as a number of other infectious agents, are believed to lose viability upon drying.

Method used

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  • Compositions and methods for the release of nucleic acid molecules from solid matrices
  • Compositions and methods for the release of nucleic acid molecules from solid matrices
  • Compositions and methods for the release of nucleic acid molecules from solid matrices

Examples

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example 1

[0164] Pine needles from individual trees (Pinus pinus) are collected at various locations in the Shenandoah National Park. Needles from each tree are placed over the surface of an FTA® CloneSaver 96 Card and crushed into the Card using a hammer. The needles are placed in the Card such that each position on the card contains sample material from only one tree. As each sample is applied to the card, a log is kept so that the sample can be linked to a particular tree. Once samples have been applied to all of the sample positions on the card, the card is stored at ambient temperature until it is transported to a lab for analysis.

[0165] Once in the lab, the samples are “punched” from the FTA® card using a HARRIS MICRO PUNCH™ Apparatus with mat. The resulting 2 mm punches are then placed in wells of a 96 well plate and washed with Tris-EDTA (pH 7.3). 50 μl of ethanolamine, adjusted to pH 11.0 with KOH, is then added to each of the wells. After incubation at 90° C. for 30 minutes, the fl...

example 2

[0166] Methods

[0167] Twenty-five μl of fresh blood was spotted onto individual locations of FTA® CloneSaver 96 Cards using a Model P-200 Pipeteman™ (Rainin Instrument Company, Inc. Rainin Road, Box 4026, Woburn, Mass., 01888). These Cards were stored at room temperature until used.

[0168] The releasing reagents were composed of ethanolamine diluted in water to a concentration of between 0.01% and 1%. The pH range was between 8.3 and 13. The pH was adjusted by the addition of aqueous KOH. The best results were obtained at a pH of 11 at a concentration of 1%. The ethanolamine reagents were compared to 15 mM and 30 mM CABS [4-(cyclohexylamino)-1-butane sulfonic acid] buffer, CAPS [3-(cyclohexylamino)-1-propane sulfonic acid] buffer, Gentra Elution Buffer 2 (used with the GENERATION™CAPTURE COLUMN™ Kit, Gentra Systems Inc., 13355 10 th Avenue N., Suite 120, Minneapolis, Minn. 55441, USA), and AmpDirect (Shimadzu, Inc., 1, Nishinokyo Kuwabaracho, Nakagyou-ku, Kyoto 604-8511, Japan).

[01...

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Abstract

The present invention relates to compositions and methods for releasing nucleic acid molecules from solid matrices. The invention further relates to compositions and methods for purifying and isolating nucleic acid molecules from biological materials such as animal tissues and plant matter. The methods of the invention can be readily adapted for rapid processing of multiple samples. Thus, the invention further provides automated methods for the purification of nucleic acid molecules from numerous samples. The invention also relates to kits for removing nucleic acid molecules from solid matrices.

Description

FIELD OF THE INVENTION [0001] The present invention relates to compositions and methods for releasing nucleic acid molecules from solid matrices. The invention further relates to compositions and methods for purifying and isolating nucleic acid molecules from biological materials such as animal tissues and plant matter. BACKGROUND OF THE INVENTION [0002] It is desirable in many instances to generate and analyze nucleic acid molecules samples obtained from numerous individual entities and / or organisms of populations. Often, such samples are used to identify the genotypes of individuals which reside either in the same or different geographic locations. Thus, the collection and analysis of samples are often employed to determine the genotypes of individual members of populations. Such analyses generally result in the generation of data relating to both the individuals from which the samples are obtained and the populations as a whole. [0003] The collection and analysis of samples which...

Claims

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Application Information

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IPC IPC(8): C12N1/15C12N1/19C12N15/09C12N1/21C12N5/10C12N15/10C12Q1/68C12Q1/6806
CPCC12N15/1006C12N15/1017C12Q1/6806C12Q2527/125
Inventor XIA, JIU-LINGOLDSBOROUGH, MINDY D.CONNOLLY, MICHAEL A.GEBEYEHU, GULILAT
Owner INVITROGEN
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