Nebnext multiplex oligos dual index primers set 1

Total RNA from radish plants and seedlings was isolated according to the modified phenol-chloroform method [111 (link)]. The cDNA was synthesized with the Mint-2 kit (Evrogen, Moscow, Russia). The NEBNext^® Ultra™ DNA Library Prep Kit for Illumina (New England Biolabs, Hitchin, UK) was used to prepare sequencing libraries. Double barcoding was performed using the NEBNext^® Ultra™ DNA Index Prep Kit for Illumina and the NEBNext^® Multiplex Oligos^® Illumina^® (Dual Index Primers Set 1). Libraries were sequenced on the Illumina HiSeq2500 sequencer.

For RNA-seq, the total RNA from the transgenic roots of radish was isolated according to the modified phenol–chloroform method [76 (link)]. For both CLE41-oe and GUS-oe genotypes, three biological samples were used. The synthesis of cDNA was performed using the Mint-2 kit (Evrogen, Moscow, Russia).
Library preparation and sequencing was performed by the Saint Petersburg State University Research Park, Centre for Molecular and Cell Technologies (Saint Petersburg, Russia). Illumina libraries were made from polyA RNA with NEBNext^® Ultra™ DNA Library Prep Kit (New England Biolabs, Hitchin, UK) according to the manual. Double barcoding was performed using the NEBNext^® Ultra™ DNA Index Prep Kit for Illumina and the NEBNext^® Multiplex Oligos^® Illumina^® (Dual Index Primers Set 1). The sequencing of libraries was performed on an Illumina HiSeq2500 sequencer with 50 bp read length.