Directional targeted sequencing

A method of analyzing a nucleic acid sample, comprising:

• (a) providing a sample comprising a plurality of nucleic acid molecules, which plurality of nucleic acid molecules comprise a plurality of target nucleic acid sequences;
• (b) subjecting said plurality of nucleic acid molecules to a condition sufficient to generate one or more copies of said plurality of target nucleic acid sequences, or complements thereof, wherein generating said one or more first copies, or complements thereof, comprises annealing first primers of a plurality of first primers and second primers of a plurality of second primers to nucleic acid molecules of said plurality of nucleic acid molecules;
• (c) separating said one or more first copies of said plurality of target nucleic acid sequences, or complements thereof, from other materials;
• and
• (d) subjecting said one or more first copies of said plurality of target nucleic acid sequences, or complements thereof, to a condition sufficient to generate one or more second copies of said plurality of target nucleic acid sequences, or complements thereof, through asymmetric PCR that anneals third primers of a plurality of third primers to said one or more first copies, or complements thereof;
• wherein said plurality of third primers are configured to anneal to said one or more first copies, or complements thereof, downstream of said first primers or second primers, or complements thereof;
• and
• wherein said plurality of third primers comprise a portion of a first sequencing adapter of said plurality of first sequencing adapters, or a complement thereof.