MPRA of Ciona robusta late gastrula whole embryos, DNA Plasmid Barcode, Replicate 1
DNA was extracted by mixing the phenol-chloroform and interphase of TriZol extraction with 500uL of Back Extraction Buffer (4M guanidine thiocyanate, 50mM sodium citrate, and 1M Tris-base). DNA was treated with RnaseA (Thermo Fisher). DNA was cleaned up with phenol:chloroform:isoamyl alcohol (25:24:1) (Life Technologies). The DNA was PCR amplified and size selected using Agencourt AMPure beads (Beckman Coulter), then checked for quality and size on the 2100 Bioanalyzer (Agilent) and sent for sequencing on the NovaSeq S4 PE100 mode (Illumina).