GSM8151818: C. elegans gonad, Worm12, Diakinesis; Caenorhabditis elegans; RNA-Seq
a cut was made across the vulva using a 26G subcutaneous needle controlled by a micromanipulator (ROE-200, Sutter) under an inverted microscope (Olympus 1X71). This would release fertilized eggs and early-stage embryos from the uterus as well as sperms and at least portions of the two sides of the gonad. Each end of the gonad wrapped around by five pairs of sheath cells was completely isolated by pushing its distal end. The distal proliferative zone (S1) was cut off around the transition zone and harvested in about 10 nl bath solution by suction using a patch clamp pipette under controlled of another micromanipulator (ROE-200, Sutter). The pachytene zone (S2), the loop corresponding to the diplotene zone (S3) and diakinesis zone (S4) were sequentially cut off and similarly harvested. The -3 (F3), -2 (F2), and -1 (F1) oocytes were also isolated by cutting through their boundaries and similarly harvested. The zygote (fertilized oocyte) also known as P0 was similarly harvested when the two pronuclei were fused at its center. We prepare a RNA-seq library for each harvested sample for Illumina platforms using a modified scRNA-seq method based on Tang et al or using the Smart-seq2 protocol