title: metatranscriptomic sequencing of BALF_NGS_2 (CRX1857571; CRA027906)
library_construction_protocol: When constructing library, DNA wiper (Vazyme #R312) was used to remove the DNA from the nucleic acid sample, and then the mixture was eluted by PCR and magnetic beads (VAHTSRNA Clean Beads, Vazyme #N412) to obtain the supernatant, and then removed the rRNA from the supernatant (Vazyme #R460). For samples with removed DNA and rRNA, double-stranded cDNA was synthesized using the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina (Vazyme #R605) kit according to the manufacturer's instructions followed by adapter ligation (VAHTSUniversal V8 RNA-seq Library Prep Kit for Illumina, Vazyme #R605). After the ligation, the library was purified using the magnetic bead (VAHTSDNA Clean Beads, Vazyme #N411), and finally the library amplification step. According to the instructions, the purified cDNA with adaptors were mixed with the reaction system mixes. After the amplification, the DNA purification with magnetic beads was repeated to complete the library construction. The constructed cDNA libraries were sent to Azenta Biotechnology Co., Ltd. (Jiangsu, China) for metatranscriptomic sequencing using the Illumina HiSeq 2000/2500 platform.