Danio rerio
Protocols: Staged embryos were transferred into 10cm dish on ice. The chorion was physically disrupted and removed by tweezers. Then the majority of the yolk was physically removed using a tweezer. The remaining embryonic cells were washed with PBS and incubated in the deyolk buffer. When single cells became dissociated, they were transferred into a new eppendorf tube and washed with PBS for three times. Nuclei acid was extracted first using a Qiagen PCR purification column. Then 1× VAHTS DNA cleaning beads were used to remove primer leftover and eluted in 20 μl Nuclease-free Water. 50,000 single cells were resuspended in 50 uL RSB + 0.01% digitonin 0.1% Tween-20 and 0.1% NP-40 and incubated on ice for 3 minutes. Then 1 mL of cold RSB + 0.1% Tween-20 (without digitonin and NP-40) was added. The permeabilised cells were centrifuged at 300 g 4 C. After removing the supernatant, 50 ul tagmentation mix was used to resuspend the cell pellet and incubated at 37C on a thermomixer for 30 minutes. Then 50 ul 20mM EDTA was added to stop the reaction. Single cells were sorted into each well of a plate containing distinct well index primers using FACS. 2x NEBNext PCR Master Mix was added into each well to perform PCR for the library construction. The PCR condition is: 72 C 5 minutes, 98 C 2 minutes, then 12 cycles of [98 C 10 seconds, 63 C 30 seconds, 72 C 20 seconds].
ENA-FIRST-PUBLIC
2023-10-14T05:00:47Z
ENA-LAST-UPDATE
2023-10-14T05:00:47Z
External Id
SAMEA113990748
INSDC center alias
Department of Systems Biology School of Life Sciences Southern University of Science And Technology
INSDC center name
Department of Systems Biology School of Life Sciences Southern University of Science And Technology
INSDC first public
2023-10-14T05:00:47Z
INSDC last update
2023-10-14T05:00:47Z
INSDC status
public
Submitter Id
E-MTAB-13115:zf_3.0hpf_plate4_335
age
3
broker name
ArrayExpress
collection date
not collected
common name
zebrafish
developmental stage
blastula 1k-cell
disease
normal
genotype
wild type genotype
geographic location (country and/or sea)
not collected
hours post fertilisation (hpf)
3
isolate
not applicable
organism part
embryonic structure
sample name
E-MTAB-13115:zf_3.0hpf_plate4_335
scientific_name
Danio rerio
sex
mixed
strain
Tubingen