Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
AssigneeBaylor College of Medicine
InventorsAllan Bradley, Wei Wen Cai
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Abstract
The invention relates to novel chemically modified nucleic acids with enhanced lability towards solid supports, such as glass. These modified nucleic acids can be readily affixed to solid supports, for instance, a glass surface, without first derivatizing the glass surface. High-density microarrays based on these modified nucleic acids as well as methods for preparing these microarrays are also useful.
Claims51
A modified nucleic acid comprising a nucleic acid covalently bound to a compound having the formula:
- R.sub.1 --X--R.sub.2
- wherein R.sub.1 is an epoxide group;
- wherein R.sub.2 is an alkoxysilane group;
- and
- wherein X is moiety, chemically suitable for linking said epoxide group and said alkoxysilane group.
A modified nucleic acid comprising a nucleic acid covalently bound to a compound having the formula:
- R.sub.1 --X--R.sub.2
- wherein R.sub.1 is an amino group;
- wherein R.sub.2 is an alkoxysilane group;
- and
- wherein X is moiety, chemically suitable for linking said amino group and said alkoxysilane group.
A device for making high-density microarrays comprising a plurality of capillary micropipets, having a distal and a tapered proximal end, wherein said micropipets are:
- arranged equatorially in a substantially close-packed arrangement;
- and
- tapered at each said proximal end and wherein said ends are essentially planar.
A high-density microarray comprising:
- a solid support;
- modified nucleic acid prepared by reacting said nucleic acid with an alkoxysilane selected from the group consisting of 3-glycidoxypropyltrimethoxysilane and 3-aminopropyltriethoxysilane, and immobilized about said solid support in orderly discrete spots.
A high-density microarray comprising a solid support having a plurality of modified silane moieties affixed, wherein said silane moieties are selected from the group consisting of --Si--(CH.sub.2).sub.n --, wherein n=3, 4, 5, 6, 7, 8, or 9;
- and --Si--CH.sub.2 --CH.sub.2 --C.sub.6 H.sub.4 --CH.sub.2 --CH.sub.2 --Si--.
A method for modulating the electrostatic properties of a glass solid support comprising selectively reacting said solid support with silanes selected from the group consisting of:
- 1,6-Bis-trichlorosilylhexane, 1,8-Bis-trichlorosilyloctane, 1,6-Bis-trimethoxysilylhexane, and 1,4-Bis-trimethyloxysilylethylbenzene.
A modified nucleic acid prepared by:
- reacting a guanine or cytosine base of said nucleic acid with N-bromosuccinimide at pH about 8.0 to form a brominated nucleic acid;
- reacting said brominated nucleic acid with a silane having the formula H.sub.2 N--(CH.sub.2).sub.n --Si(OC.sub.2 H.sub.5).sub.3, wherein n=3, 4, 5, 6, 7, 8, or 9.
A modified nucleic acid comprising a nucleic acid covalently bound to a compound having the formula:
- --HN--(CH.sub.2).sub.n --Si(OR).sub.3
- wherein n=3, 4, 5, 6, 7, 8, or 9.
A modified nucleic acid comprising a nucleic acid covalently bonded to a compound having the formula:
- ##STR4## wherein R is selected from the group consisting of --CH.sub.3, --C.sub.2 H.sub.5, and --C.sub.3 H.sub.7 ;
- wherein R.sub.1 and R.sub.2 are the same or different and are selected from the group consisting of --H, --CH.sub.3, --C.sub.2 H.sub.5, --OCH.sub.3, --OC.sub.2 H.sub.5, --C.sub.3 H.sub.7, and --OC.sub.3 H.sub.7 ;
- and
- wherein X linking group comprising an at least partially aliphatic chain.
A method for immobilizing nucleic acid to solid support comprising:
- reacting a compound of the formula R.sub.1 --X--R.sub.2 with a nucleic acid to form a derivatized nucleic acid;
- wherein R.sub.1 is an epoxide group;
- wherein R.sub.2 is an alkoxysilane group;
- and, wherein X is a moiety chemically suitable for linking said epoxide group and said alkoxysilane group;
- and,
- reacting said derivatized nucleic acid with said solid support.
A method for immobilizing nucleic acid to solid support comprising:
- reacting compound of the formula R.sub.1 --X--R.sub.2 with a nucleic acid to form a derivatized nucleic acid;
- wherein R.sub.1 is an amino group;
- wherein R.sub.2 is an alkoxysilane group;
- and, wherein X is a moiety chemically suitable for linking said amino group and said alkoxysilane group;
- and
- reacting said derivatized nucleic acid with said solid support.
A modified nucleic acid comprising a nucleic acid covalently bound to a compound having the formula:
- R.sub.1 --X--R.sub.2
- wherein R.sub.1 is a cyclic ether;
- wherein R.sub.2 is --NR.sub.4, wherein each R.sub.3 group are the same or different alkyl groups, and R.sub.4 is --H or an alkyl group;
- and
- wherein X is moiety, chemically suitable for linking said cyclic ether group and said alkoxysilane group.
A modified nucleic acid comprising a nucleic acid covalently boded to a compound having the formula:
- ##STR5## wherein R.sub.1, R.sub.2, and R.sub.3, are the same or different, and are selected from the group consisting of --OCH.sub.3, --OC.sub.2 H.sub.5, --OC.sub.3 H.sub.7, and --Cl;
- and
- wherein X is a moiety, chemically suitable for linking said nucleic acid to said compound.
Dependencies are inferred from the claim text.