RNA-Seq of K. marxianus 3444 strain 2: LBA production
The library was constructed using the Illumina TruSeqTM RNA sample prep Kit. Initially, polyadenylated eukaryotic mRNA was enriched using magnetic beads coated with Oligo(dT), followed by fragmentation of the mRNA using sonication. Fragmented mRNA was then used as a template, with random oligonucleotides serving as primers, for the synthesis of the first cDNA strand in the presence of M-MuLV reverse transcriptase. Subsequently, the RNA strand was degraded by RNaseH, and the second cDNA strand was synthesized using dNTPs as substrates in the presence of DNA polymerase I. The purified double-stranded cDNA underwent end repair, A-tailing, and ligation of sequencing adapters. The cDNA fragments of approximately 200bp were size-selected using AMPure XP beads, followed by PCR amplification and purification of the PCR products using AMPure XP beads. Finally, the library was obtained.