GSM8658801: rapgef1b KD embryo, 1 dpf_n3; Danio rerio; RNA-Seq
The total RNA was extracted from 1dpf control and rapgef1b depleted embryos using an RNA isolation kit (MN; 740955.50). Sequencing libraries were generated using MGIEasy RNA library prep set (MGI) according to the manufacturer's instructions. Here, 500 ng total RNA was used as starting material, and rRNA was depleted by MGIEasy rRNA depletion Kit. The rRNA-depleted samples were fragmented, reverse transcribed and the second strand was synthesized and converted to cDNA. DNA was then purified using DNA Clean Beads provided in the kit followed by end repair and A-tailing. Barcoding and adaptor ligation were performed and the samples were purified. Samples were amplified using adaptor-specific primers and quantified using a Qubit dsDNA high-sensitivity kit (ThermoScientific). Sample fragment size was determined using 4200 TapeStation (Agilent). 1pMol of the dsDNA library was denatured and circularized to make single-stranded circular DNA. DNA Nano Balls (DNBs) were created using Rolling cycle amplification. DNBs were loaded onto patterned flowcell and sequenced using PE100 recipe on MGISEQ-2000 sequencer (MGI). Barcoding and adaptor ligation were performed and the samples were purified. Samples were amplified using adaptor-specific primers and quantified using a Qubit dsDNA high-sensitivity kit (ThermoScientific). Sample fragment size was determined using 4200 TapeStation (Agilent). 1pMol of the dsDNA library was denatured and circularized to make single-stranded circular DNA. DNA Nano Balls (DNBs) were created using Rolling cycle amplification. DNBs were loaded onto patterned flowcell and sequenced using PE100 recipe on MGISEQ-2000 sequencer (MGI).
GEO
GSM8658801_r1