Compositions and Methods for Detecting C1orf43 Nucleic Acid
This disclosure provides oligomers, combinations of oligomers, compositions, kits, uses, and methods for detecting a C1orf43 nucleic acid, such as C1orf43 mRNA, such as human C1orf43 mRNA, in a sample.
1 . A kit comprising:
(a) a combination of oligomers comprising at least first and second amplification oligomers, wherein the first and second amplification oligomers are reverse and forward amplification oligomers, respectively, and are configured to specifically hybridize to first and second sites in the sequence of SEQ ID NO: 39 and generate an amplicon therefrom, respectively;
wherein the first amplification oligomer comprises a target-hybridizing sequence that is from 14 to 50 bases in length and comprises the sequence of SEQ ID NO: 53 or SEQ ID NO: 54;
wherein the second amplification oligomer comprises a target-hybridizing sequence that is from 15 to 40 bases in length and comprises the sequence of SEQ ID NO: 47 or SEQ ID NO: 48; and
wherein at least one of the amplification oligomers is a promoter-primer and/or the combination further comprises at least one probe oligomer that comprises at least 10 nucleotides, is configured to specifically hybridize to an amplicon produced from the first and second amplification oligomers, and (i) comprises 2′-O-methyl-ribose in its backbone and/or (ii) comprises a non-nucleotide detectable label; and
(b) an enzyme.
2 . The kit of claim 1 , wherein at least one of the amplification oligomers is a promoter-primer.
3 . The kit of claim 1 , wherein the first amplification oligomer is a promoter primer.
4 . The kit of claim 3 , wherein the promoter-primer comprises a T7 promoter which is located 5′ of the target-hybridizing sequence and wherein the T7 promoter comprises the sequence of SEQ ID NO: 58.
5 . The kit of claim 1 , wherein the target-hybridizing sequence of the first amplification oligomer comprises the sequence of SEQ ID NO: 27, 29, 31, or 33.
6 . The kit of claim 1 , wherein the first amplification oligomer comprises the sequence of SEQ ID NO: 26, 28, 30, or 32.
7 . The kit of claim 1 , wherein the target-hybridizing sequence of the second amplification oligomer comprises the sequence of SEQ ID NO: 34, 35, 36, 37, or 38.
8 . The kit of claim 1 , wherein the combination further comprises at least one probe oligomer that comprises at least 10 nucleotides and is configured to specifically hybridize to an amplicon produced from the first and second amplification oligomers.
9 . The kit of claim 8 , wherein the probe oligomer is configured to specifically hybridize to a detection site in a nucleic acid having the sequence of SEQ ID NO: 49.
10 . The kit of claim 8 , wherein the probe oligomer comprises a target hybridizing sequence comprising the sequence of SEQ ID NO: 15, 17, 19, 21, 23, or 25.
11 . The kit of claim 8 , wherein the probe oligomer comprises the sequence of SEQ ID NO: 14, 16, 18, 20, 22, or 24.
12 . The kit of claim 8 , wherein the probe oligomer comprises 2′-O-methyl-ribose in its backbone.
13 . The kit of claim 8 , wherein the probe oligomer comprises a non-nucleotide detectable label.
14 . The kit of claim 13 , wherein the non-nucleotide detectable label is a fluorescent label.
15 . The kit of claim 14 , wherein the probe oligomer comprise a quencher.
16 . The kit of claim 13 , wherein the non-nucleotide detectable label is a chemiluminescent label.
17 . The kit of claim 1 , wherein the enzyme is a reverse transcriptase.
18 . The kit of claim 1 , wherein the enzyme is an RNA polymerase.
19 . The kit of claim 17 , further comprising a second enzyme, wherein the second enzyme is an RNA polymerase.
20 . The kit of claim 1 , wherein the combination of oligomers is contained in a lyophilized composition.