Accelerated nucleic acid reassociation method

Claim 1

An improved method for the formation of double-stranded nucleic acid molecules from separate single-stranded nucleic acid molecules wherein the rate of reaction is greatly increased over the standard reference condition reaction rate, said method comprising the steps of: preparing an aqueous reaction solution containing a quantity of a first single-stranded nucleic acid molecule and a quantity of a second single-stranded nucleic acid molecule, said second single-stranded nucleic acid molecule having at least one segment of base sequences complementary to a corresponding segment of base sequences of said first single-stranded nucleic acid molecule, and a known concentration of at least one nucleic acid precipitating agent, said known concentration being sufficient to greatly accelerate the rate of reaction over the rate of the standard reference condition reaction; incubating said aqueous reaction solution at a temperature at which reassociation can occur; and assaying said incubated aqueous reaction solution for the presence of double-stranded nucleic acid molecules.

Claim 5

The method of any one of the preceding claims wherein said aqueous reaction solution also contains a known concentration of a nucleic acid denaturing agent.

Claim 8

The method of any one of the preceding claims wherein the pH of said aqueous reaction solution ranges from about 4 to about 11.

Claim 9

The method of any one of the preceding claims wherein said temperature ranges from about 20°C to about 90°C.

Claim 10

The method of any one of the preceding claims wherein said strong nucleic acid precipitating agent is sodium phosphate.

Claim 11

The method of any one of claims 1 to 8 wherein said nucleic acid precipitating agent is NaS0 4 .

Claim 12

The method of any one of claims 1 to 8 wherein said nucleic acid precipitating agent is LiCl.

Claim 13

The method of any one of claims 1 to 8 wherein said nucleic acid precipitating agent is (NH 4 ) 2 SO 4 .

Claim 14

The method of any one of claims 1 to 8 wherein said nucleic acid precipitating agent is 13 volume % to 30 volume % Sarkosyl.

Claim 15

The method of any one of the preceding claims wherein said nucleic acid precipitating agent is one miscible with an aqueous solution and capable of precipitating single-stranded nucleic acid molecules from an aqueous solution.

Claim 17

The method according to any one of the preceding claims comprising: screening a suspected nucleic acid precipitating agent to determine the effective concentration of said suspected nucleic acid precipitating agent and to determine the associated incubation temperature necessary to greatly increase the rate of reaction over the standard reference condition reaction rate; and incubating said aqueous reaction solution including said effective concentration of said nucleic acid precipitating agent.

Claim 19

A method for screening a suspected nucleic acid precipitating agent for use in an accelerated nucleic acid reassociation reaction, said method comprising the steps of: preparing a series of solutions, each of said solutions containing a known amount of single-stranded nucleic acid and a different amount of said suspected nucleic acid precipitating agent; examining each of said solutions for the presence of a nucleic acid precipitate; and heating each of said solutions in which a precipitate was found in order to determine whether the degree of precipitation changes.

Claim 20

A method for screening a suspected nucleic acid precipitating agent for use in an accelerated nucleic acid reassociation reaction, said method comprising the steps of: preparing a series of solutions, each of said solutions containing a known amount of single-stranded nucleic acid and a different amount of said suspected nucleic acid precipitating agent; examining each of said solutions for the presence of a nucleic acid precipitate; and heating each of said solutions in which a precipitate was found in order to determine whether the degree of precipitation changes.