23 ug of starting sample was taken for Sonication on Covaris S220 (as per the SOLiD Mate Pair Library prep protocol), analyzed for size distribution post-sonication (on Agilent Bioanalyzer High Sensitivity chip). Fragments were size selected at the range of 2.5-3.5kb on 0.6 % Agarose, and size distribution was verified on 2% E-Gel. Following this the fragments were end-repaired and MPR-MPL adaptor ligated. Nick-translation was performed on circularized adaptor ligated DNA, and digested with T7 followed by S1 Nuclease enzymes and 3’ Adenylation by P1-T and P2-T. Post ligation the sample was cleaned using Streptavidin beads. Adaptor ligated sample was amplified with 18 cycles of PCR and size selected (range of 250bp to 350bp using E-Gel), followed by analysis on Bioanalyzer High Sensitivity chip for assessing library size distribution. Library fragments were amplified on beads by emulsion PCR. One single sequence read that covers the two original ends and the internal adapter is generated. Sequencing was performed with two different primers, complementary to the P1 adapter and internal adapter, respectively. The resulting read pairs occur in same direction and orientation.

Bioproject ID: PRJNA267195, Accession ID: SRX761338, Run ID: SRR1654829, Experiment ID: SRX761338, Biosample ID: SAMN03198752 (SRS745872) (Ref.: SRA, NCBI, USA)

O. sanctum (CIM-Ayu) 4 months old leaf (Syn. Krishna Tulsi)