Patent ID: 8278035

Claim:
A method for isolating a microRNA of interest ( 42 ) from a sample comprising the microRNA of interest ( 42 ) according to FIG. 1 through FIG. 6 ; the method comprising: a) providing a sample comprising the microRNA of interest ( 42 ); b) providing a capture probe ( 10 ) comprising: i) a first adapter segment ( 12 ) having a first adapter segment sequence, the first adapter segment comprising a 3′ end ( 18 ) and a 5′ end ( 20 ); ii) a second adapter segment ( 14 ) having a second adapter segment sequence, the second adapter segment comprising a 3′ end ( 30 ) and a 5′ end ( 32 ); and iii) a microRNA binding segment ( 16 ) having a microRNA binding segment sequence; where the microRNA binding segment ( 16 ) is substantially complementary to, and capable of hybridizing to, one or more than one microRNA of interest ( 42 ) by Watson-Crick base pairing; where the 5′ end of the first adapter segment ( 20 ) is connected to the 3′ end of the microRNA binding segment ( 22 ); and where the 3′ end of the second adapter segment ( 30 ) is connected to the 5′ end of the microRNA binding segment ( 32 ); c) providing a first linker ( 48 ), wherein the first linker has a first linker sequence, comprises a 3′ end ( 52 ) and a 5′ end ( 54 ), and is substantially complementary to, and capable of hybridizing to, the first adapter segment ( 12 ) of the capture probe ( 10 ); d) providing a second linker ( 50 ), wherein the second linker has a second linker sequence, comprises a 3′ end ( 56 ) and a 5′ end ( 58 ), and is substantially complementary to, and capable of hybridizing to, the second adapter segment ( 14 ) of the capture probe ( 10 ); e) combining the sample, the capture probe ( 10 ), the first linker ( 48 ) and the second linker ( 50 ) in a solution; f) allowing the first linker ( 48 ) to hybridize with the first adapter segment ( 12 ), the microRNA of interest ( 42 ) to hybridize with the microRNA binding segment ( 16 ), and the second linker ( 50 ) to hybridize with the second adapter segment ( 14 ); g) ligating the 3′ end of the first linker ( 52 ) to the 5′ end of the microRNA of interest ( 46 ), and ligating the 3′ end of the microRNA of interest ( 44 ) to the 5′ end of the second linker ( 58 ), thereby producing a complex defined as a strand of first linker, microRNA of interest and second linker that have been ligated together (ligated first linker ( 48 )-microRNA of interest ( 42 )-second linker ( 50 )) ( 62 ) and that is hybridized to the capture probe ( 10 ); h) dehybridizing the capture probe ( 10 ) from the strand of the ligated first linker-microRNA of interest-second linker ( 62 ); and i) purifying the ligated first linker-microRNA of interest-second linker ( 62 ), thereby purifying the microRNA of interest ( 42 ).