RNA-seq of Phrynus marginemaculatus: leg 4
TruSeq Stranded mRNA Sample Preparation Guide Rev. E; Stranded rgts removed from -20C and thawed according to protocol directions; Protocol: (19); 200ng total RNA input (96-well plate); Poly-A selection; Fragment, Prime, Finish using FPF mix (Frag 5 minutes); First strand cDNA Synthesis; Second strand cDNA Synthesis; Agencourt AMPure XP beads bead purification (lot 16572300); Plate stored at -20C; 7/16/18; Plate removed from -20C, thawed on ice and quick spun; Adenylate 3 Ends; Ligation- Dual CD Index (CD plate lot# 20105027); Agencourt AMPure XP beads bead purification (lot 16572300) x2; PCR (15 cycles) - plate left in thermal cycler at 4C ON; 7/17/18; Plate removed from thermal cycler followed by quick spin; 2 TruSeq Stranded mRNA Sample Preparation Guide Rev. E; Agencourt AMPure XP 0.8X bead purification (lot# 16572300); Added 32.5ul RSB & recovered 30ul of purified library; Qubit Quantification- 1:100 dilution (2ul) of library; Qubit HS DNA Rgt Lot# 1941055; Libraries submitted in 96-well plate to DNA Sequencing Facility for final Agilent QC and sequencing; University of Wisconsin Biotechnology Center; Gene Expression Center; 425 Henry Mall; Madison, WI 53706; www.biotech.wisc.edu/gec