A method for preparing an RNA sample for a target specific next generation sequencing comprising performing a one-step target enrichment in a single reaction vessel or in a single reaction mixture, wherein said enrichment comprises the steps: (i) exposing the RNA to be sequenced in a single reaction vessel to a mixture comprising a reverse transcriptase, a DNA polymerase, and (a) one or more target-specific reverse primers, suitable for the preparation of a target specific cDNA; and (b) a forward target specific primer comprising a first forward adaptor sequence, and a reverse target specific primer comprising a first reverse adaptor sequence, and (c) a forward indexing primer comprising a first forward adaptor sequence, a forward indexing sequence and a second forward adaptor sequence; and a reverse indexing primer, comprising a first reverse adaptor sequence, a reverse indexing sequence, wherein the reverse indexing sequence is different from the forward indexing sequence, and a second reverse adaptor sequence; and desoxyribonucleoside triphosphates (dNTPs); and (ii) subjecting the reaction mixture of (i) to a series of temperature changes under conditions sufficient to yield a first strand cDNA copy of at least a portion of the RNA to be sequenced, preferably a gene sequence, and subsequently a target specific amplicon comprising starting from the 5'- to the 3'-end a second forward adaptor sequence, a forward indexing sequence, a first forward adaptor sequence, a forward target specific primer sequence, target sequence, a reverse target specific primer sequence, a first reverse adaptor sequence, a reverse indexing sequence and a second reverse adaptor sequence. (i) exposing the RNA to be sequenced in a single reaction vessel to a mixture comprising a reverse transcriptase, a DNA polymerase, and (a) one or more target-specific reverse primers, suitable for the preparation of a target specific cDNA; and (b) a forward target specific primer comprising a first forward adaptor sequence, and a reverse target specific primer comprising a first reverse adaptor sequence, and (c) a forward indexing primer comprising a first forward adaptor sequence, a forward indexing sequence and a second forward adaptor sequence; and a reverse indexing primer, comprising a first reverse adaptor sequence, a reverse indexing sequence, wherein the reverse indexing sequence is different from the forward indexing sequence, and a second reverse adaptor sequence; and desoxyribonucleoside triphosphates (dNTPs); and (a) one or more target-specific reverse primers, suitable for the preparation of a target specific cDNA; and (b) a forward target specific primer comprising a first forward adaptor sequence, and a reverse target specific primer comprising a first reverse adaptor sequence, and (c) a forward indexing primer comprising a first forward adaptor sequence, a forward indexing sequence and a second forward adaptor sequence; and a reverse indexing primer, comprising a first reverse adaptor sequence, a reverse indexing sequence, wherein the reverse indexing sequence is different from the forward indexing sequence, and a second reverse adaptor sequence; and desoxyribonucleoside triphosphates (dNTPs); and (ii) subjecting the reaction mixture of (i) to a series of temperature changes under conditions sufficient to yield a first strand cDNA copy of at least a portion of the RNA to be sequenced, preferably a gene sequence, and subsequently a target specific amplicon comprising starting from the 5'- to the 3'-end a second forward adaptor sequence, a forward indexing sequence, a first forward adaptor sequence, a forward target specific primer sequence, target sequence, a reverse target specific primer sequence, a first reverse adaptor sequence, a reverse indexing sequence and a second reverse adaptor sequence. <EOS>