Methods and kits for room temperature in situ detection of a target nucleic acid in a biological sample

摘要

The present invention relates to in situ hybridization methods for detecting a target nucleic acid in a biological sample comprising performing one or more method steps (e.g., pretreatment, denaturation, hybridization, washes) at room temperature. The invention further relates to kits for performing such methods.

主权项

1. A kit for detecting a target nucleic acid in a biological sample, comprising:
a) a denaturation buffer comprising about 0.03M to about 0.17M base and about 50% to about 80% alcohol; b) a hybridization buffer comprising about 1-10 mM base and having a pH in the range from about 10 to about 13, wherein the base is potassium hydroxide, barium hydroxide, caesium hydroxide, sodium hydroxide, strontium hydroxide, calcium hydroxide, lithium hydroxide, rubidium hydroxide, magnesium hydroxide, butyl lithium, lithium diisopropylamide, lithium diethylamide, sodium amide, sodium hydride, lithium bis(trimethylsilyl)amide, sodium carbonate or ammonia, or a combination thereof; and c) a wash buffer comprising about 1-10 mM base, wherein the wash buffer has a pH in the range from about 10 to about 13, wherein the base is potassium hydroxide, barium hydroxide, caesium hydroxide, sodium hydroxide, strontium hydroxide, calcium hydroxide, lithium hydroxide, rubidium hydroxide, magnesium hydroxide, butyl lithium, lithium diisopropylamide, lithium diethylamide, sodium amide, sodium hydride, lithium bis(trimethylsilyl)amide, sodium carbonate or ammonia, or a combination thereof.