Method for nucleic acid detection by guiding through a nanopore

A panel of two or more guide RNAs (gRNAs), each gRNA comprising:

• (i) a nucleotide sequence that binds to a sequence in the target polynucleotide,
• (ii) a nucleotide sequence that binds to a polynucleotide guided effector protein, and
• (iii) an adaptor and/or an anchor capable of coupling to a surface,
• wherein each guide RNA comprises a crRNA and a tracrRNA, wherein the anchor or the adaptor is present at the 5′ end of the tracrRNA, the 3′ end of the tracrRNA, the 3′ end of the crRNA, or internally wherein the tracrRNA and crRNA are comprised in a sgRNA, wherein the two or more guide RNAs bind to different sequences in the same target polynucleotide.

A panel of two or more guide RNAs, each guide RNA comprising:

• (i) a nucleotide sequence that binds to a sequence in a target polynucleotide;
• (ii) a nucleotide sequence that binds to a polynucleotide guided effector protein;
• and
• (iii) an anchor capable of coupling to a surface or an adaptor, wherein each guide RNA comprises a crRNA and a tracrRNA,
• wherein the anchor or the adaptor is present at the 5′ end of the tracrRNA, the 3′ end of the tracrRNA, the 3′ end of the crRNA, or internally wherein the tracrRNA and crRNA are comprised in a sgRNA, wherein the two or more guide RNAs bind to different sequences in the same target polynucleotide, wherein the adaptor further comprises a polynucleotide binding protein capable of moving along a polynucleotide attached thereto and the anchor comprises cholesterol.