organism: Arabidopsis thaliana
description: Protocols: Arabidopsis thaliana stage 9 flower buds were collected and squeezed between a glass slide and coverslip to release meiocytes in prophase I (Walker et al, 2018 Nat. Genet.). Clean meiocytes free from somatic-cell debris were transferred to a new slide, washed with 1×PBS buffer and frozen in liquid nitrogen. ~2,000 meiotic cells were collected for each replicate. Arabidopsis thaliana plants were grown under long day 16/8 hour light/dark photoperiods in a growth chamber at 21 °C and 70% humidity. Cells were lysed with lysis buffer (0.2% triton-X100 with RNase inhibitor), followed by reverse transcription using oligo-dT30VN (Sigma-Aldrich) and Superscript IV (Thermofisher). cDNA was amplified using KAPA HiFi HotStart ReadyMix (ROCHE DIAGNOSTICS LTD) for 14 cycles before purification using Agencourt AMPure beads (Beckman Coulter). Pre-amplified libraries were quantified and quality controlled by Bioanlayzer using the High Sensitivity DNA Kit (Agilent, US). PCR amplicons were constructed into dual-indexed libraries using a Nextera XT kit (Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-01-03T15:59:19Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-04-15T14:21:11Z
attributes.tag: External Id
attributes.value: SAMEA5567477
attributes.tag: INSDC center name
attributes.value: UNIVERSITY OF CAMBRIDGE
attributes.tag: INSDC first public
attributes.value: 2020-01-03T15:59:19Z
attributes.tag: INSDC last update
attributes.value: 2019-04-15T14:21:11Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7870:Bur_Rep1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: sporocyte
attributes.tag: common name
attributes.value: thale cress
attributes.tag: developmental stage
attributes.value: sporophyte senescent stage
attributes.tag: ecotype
attributes.value: Bur-0
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: growth condition
attributes.value: 16 hours light; 21C
attributes.tag: organism part
attributes.value: flower bud
attributes.tag: sample name
attributes.value: E-MTAB-7870:Bur_Rep1
attributes.tag: scientific_name
attributes.value: Arabidopsis thaliana