lib3
TRF-1 control
Illumina-compatible PolyA enriched RNA sequencing library preparation for Tea RNA was performed at Genotypic Technology Pvt. Ltd. Genomics facility. Transcriptome library for sequencing was constructed according to NEBNext Ultra Directional Library Prep Kit (NEB, #E7420S). 200ng of Qubit quantified Total RNA was taken for mRNA isolation, fragmentation and priming. The fragmented and primed mRNA was further subjected to first and second strand synthesis. The double stranded cDNA was cleaned up using HighPrep PCR (Magbio, Cat #AC-60050) beads. Subsequently, cDNA was subjected to End Repair, A Tailing followed by adaptor ligation. The adaptor ligated cDNA was cleaned up using HighPrep PCR beads and was followed by 15 cycles of Indexing-PCR (37 C for 15mins followed by denaturation at 98C for 30 sec, cycling (98C for 10sec, 65C for 75sec) and 65C for 5mins) to enrich the adaptor-ligated fragments. The PCR product was purified using HighPrep beads. Quantification and size distribution of the prepared libraries were determined using Qubit flourometer and the Agilent D1000 TapeStation respectively according to the manufacturers instructions . The adapter ligated molecules were quantified on qPCR using KAPA library quantification kit (KAPA Biosystems).
lib3
SUB5569172
lib3
Authors name list
1. M. Nalina, Division of Plant Physiology and Biotechnology, UPASI Tea Research Foundation, Tea Research Institute, Nirar Dam, Valparai 642 127, Coimbatore, Tamil Nadu, India 2. M. Prakash Navale, Division of Biotechnology, Indian Institute of Horticultural Research, Hessarghatta Lake Post, Bangalore 560 089, Karnataka, India 3. Mohamed Aiyaz, Genotypic Technology Pvt Ltd., #2/13, Balaji Complex, R.M.V. 2nd Stage, Bangalore 560 094, Karnataka, India 4. K. N. Chandrashekara, Division of Plant Physiology and Biotechnology, UPASI Tea Research Foundation, Tea Research Institute, Nirar Dam, Valparai 642 127, Coimbatore, Tamil Nadu, India