GSM8678136: RfxCas13d control, rep2; Danio rerio; RNA-Seq
Total RNA then was extracted using Direct-zol RNA Miniprep Kit (#R2050, Zymo Research) following manufacturer's instructions. Zebrafish embryos total RNA sequencing libraries were generated from 100 ng of high-quality total RNA, as assessed using the Bioanalyzer (Agilent), with the Watchmaker mRNA Library Prep Kit (Watchmaker, Cat. No. 7BK0001) at a 1/5th reaction volume. mRNA isolation and library construction were completed per protocol at the miniaturization scale. For Library amplification, 2 µL of IDT indexed primers (using xGen™ Stubby Adapter-UDI Primers, 96 reactions, Cat No. 10005921) was added to 3 µL of library, and 5 µL of Equinox Amplification Master Mix (2x). These libraries were then amplified for 12 cycles of PCR using an annealing temperature of 55°C according to step 10.2. 1.0x SPRISelect (Beckman Coulter, Cat. No. B23318) bead ratio was used during the Post-Amplification Cleanup, and post amplification libraries were eluted in 10 µL of Qiagen EB (Qiagen, Cat. No. 19086). Resulting short fragment libraries were checked for quality and quantity using the Bioanalyzer and the Qubit Flex Fluorometer (Life Technologies). Equal molar libraries were pooled, quantified, and converted to process on the Element AVITI.