title: GSM6542035: Hypsibius exemplaris active biol rep 3; Hypsibius exemplaris; RNA-Seq
library_construction_protocol: Specimens were thoroughly washed with Milli-Q water on a sterile nylon mesh (Millipore), and were placed in a low-binding PCR tube. For active individuals, pipette tips were used to homogenize the animals and lysed with the TRIzol reagent (Life Technologies). Total RNA was extracted using a Direct-zol RNA kit (Zymo Research), and the quality was validated with High Sensitivity RNA ScreenTape on TapeStation 2200 (Agilent Technologies). For storage cells, collected storage cells were directly put into the lysis buffer of SMART-Seq v.4 kit (Clontech). The mRNA was amplified with the SMART-Seq v.4, and Illumina libraries were prepared with the KAPA HyperPlus Kit (KAPA Biosystems). The library was quantified using the Qubit Fluorometer (Life Technologies), and size distribution was validated with TapeStation D1000 ScreenTape (Agilent Technologies). The library was size-selected for 200 bp by manually cutting out an agarose gel and was purified with a NucleoSpin Gel and PCR Clean-up kit (Clontech).