Becton Dickinson and Company Limited

Adriann J Howard

Daniel L Woodard

James A Down

William E Keating

Jillian A Robson

The present invention relates to a silicon-containing material which exhibits sufficient hydrophilicity and sufficient electropositivity to bind DNA from a suspension containing DNA and permit elution of the DNA from the material. Generally, the hydrophilic and electropositive characteristics are expressed at the surface of the silicon-containing material. Preferred silicon-containing materials of the present invention include boron silicate, aluminum silicate, phosphosilicate, silica carbonyl, silica sulfonyl and silica phosphonyl. The silicon-containing materials of the present invention are particularly useful in processes for purification of DNA from other cellular components. In these processes, a suspension of cellular components is placed in contact with the silicon-containing material, the silicon-containing material is washed to remove all cellular components other than DNA which are bound to the material, and the bound DNA is eluted from the material. Several of the silicon-containing materials are capable of binding and eluting DNA using only water.

DNA purification on modified siligates

The present invention relates to modified glass fiber membranes which exhibit sufficient hydrophilicity and sufficient electropositivity to bind DNA from a suspension containing DNA and permit elution of the DNA from the membrane. Generally, the hydrophilic and electropositive characteristics are expressed at the surface of the modified glass fiber membrane. Preferred modified glass fiber membranes of the present invention include glass fiber membranes that have been modified by treatment with trifluoroacetic acid (TFA), BCl.sub.3, SiCl.sub.4, NaOH, F.sup.-, AlCl.sub.3 alone or in combination, with or without water. The modified glass fiber membranes of the present invention are particularly useful in processes for purification of DNA from other cellular components. In these processes, a suspension of cellular components is placed in contact with the modified glass fiber membrane, the modified glass fiber membrane is washed to remove all cellular components other than DNA which are bound to the membrane, and the bound DNA is eluted from the membrane.

Modified glass fiber membranes useful for DNA purification by solid

The present invention relates to fluorinated surfaces which exhibit sufficient hydrophilicity and sufficient electropositivity to bind DNA from a suspension containing DNA and permit elution of the DNA from the surface. Generally, the hydrophilic and electropositive characteristics are expressed at the fluorinated surface. Preferred fluorinated surfaces of the present invention include fluorinated Al(OH).sub.3, fluorinated SiO.sub.2 and fluorinated Celite. The fluorinated surfaces of the present invention are particularly useful in processes for purification of DNA from other cellular components. In these processes, a suspension of cellular components is placed in contact with the fluorinated surface, the fluorinated surface is washed to remove all cellular components other than DNA which are bound to the surface, and the bound DNA is eluted from the surface. Lower concentrations of chaotrope in the binding buffer are needed to bind DNA to the fluorinated surfaces.

DNA purification by solid phase extraction using fluorinated celite

DNA purification by solid phase extraction using a hydroxide-washed

DNA purification by solid phase extraction using glass fiber membrane

Glass fiber membranes modified by treatment with SiCl.sub.4, AlCl.sub.3

DNA purification by solid phase extraction using trifluorometric acid

Silicon tetrahydrazide compounds produced by reacting SiCl.sub.4 with hydrazine to completion. The compounds are useful for binding DNA, as they do not require the use of chaotrope or alcohol solutions for binding. Binding and elution of DNA can be accomplished in low salt buffers or water. Kits containing the compounds for purification of DNA are also disclosed.

Silicon tetrahydrazide for purification of DNA

Boron silicates, phosphosilicates and aluminum silicates useful as binding surfaces for DNA purification. These compounds allow DNA to be bound and eluted under native conditions (i.e., in the absence of chaotropes or alcohols) using only water, low salt buffers or physiological buffers. Methods for preparation of the compounds and methods for purifying DNA using the compounds are also provided.

Boron silicates, aluminum silicates, and phosphosilicates for

A process for purifying DNA comprising binding the DNA to a silicate compound in the presence of water, low salt buffer, or physiological buffer and eluting in the bound DNA by heating in a low salt buffer, a physiological buffer, or water. The silicate compound is prepared by reacting SiCl.sub.4 with a molar percentage of BCl.sub.3 of about 33.5%, 55.6%, or 83.3%, with a molar percentage of PCl.sub.3 of about 50.0%-83.3%, or with a molar percentage of AlCl.sub.3 of about 9.1%; cooling the reaction mixture; adding water until the evolution of gas is complete; and recovering the compound.

Purification of DNA on aluminum silicates, and phosphosilicates

A process for preparing borosilicate, aluminosilicate, or phosphosilicate comprising preparing a mixture of SiCl.sub.4 with a specific molar ratio of BCl.sub.3, AlCl.sub.3, or PCl.sub.3 ; cooling said mixture to zero degrees; and adding water to the reaction mixture until the evolution of gas is complete. These silicates are useful for purifying DNA wherein the DNA is bound in water or low salt buffers (i.e., in the absence of a chaotropic agent or alcohol).

Process for preparing boron, aluminum, and phosphorus silicates for

Silicate compounds for DNA purification

DNA purification by solid phase extraction using a PCl.sub.3 modified

DNA purification by solid phase extraction using partially fluorinated

The invention provides a rapid process for lysing Mycobacteria. In one embodiment is provided a process for lysing Mycobacteria which comprises exposing the bacteria to a lysis effective amount of heat. The process of the invention is particularly advantageous since only one step is involved, it is expedient compared to prior methods, and little instrumentation is necessary. By practicing the present invention it is possible to lyse Mycobacteria with minimal effort. In addition, practicing the invention results in liberating cellular components including deoxyribonucleic acid (DNA) from Mycobacteria. Not only is DNA liberated, but the DNA is suited for subsequent analysis by way of probe hybridization, restriction enzyme analysis, and the like.

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Raleigh, NC, United States of America

Adriann J Howard