vahts dna clean beads  (Vazyme Biotech Co)

Journal: PLOS Pathogens

Article Title: Large-scale genome re-annotation uncovers pathogen-specific lncRNA regulators of fungal growth and virulence

doi: 10.1371/journal.ppat.1014681

Figure Lengend Snippet: (A) PCR amplification of genomic DNA and cDNA for ssp7 and ssp8 using primers spanning their full-length transcripts. Manually curated transcript sizes are indicated, and experimentally obtained cDNA bands are marked with red circles. tubulin served as a control. (B) Schematic comparison of predicted and actual exon-intron structures for ssp7 and ssp8 . (C) Relative mRNA expression of ssp7 and ssp8 in the wild-type strain cultured in minimal medium (MM) and mouse lung-homogenate medium (Mice Lung), quantified by RT-qPCR. Statistical significance was determined using unpaired t test (n ≥ 4). **** P < 0.0001. (D) Volcano plot of proteomic comparison of the wild-type strain cultured in MM (control) and Mice Lung (treatment). (E) Western blot analysis of FLAG-tagged Ssp7- and Ssp8-expressing strains. Actin served as a loading control. (F, H) Predicted ORFs within the ssp7 / ssp8 genomic locus. Four predicted ORFs and their reading frames are shown with start and stop positions. Point mutations (ATG → AAG) introduced into each predicted ORF are indicated. (G, I) Colony morphology and diameters of related strains grown on solid MM medium at 37°C for 48 h. Representative images are shown alongside quantification of colony diameters (n = 6). Statistical analysis was performed using one-way ANOVA with multiple comparisons tests. **** P < 0.0001; ns, not significant.

Article Snippet: The PCR products were purified using 0.9 × DNA Clean Beads (N411, Vazyme), and library amplification was performed using the VAHTS RNA Multiplex Oligos Set1 for Illumina kit (N323, Vazyme).

Techniques: Amplification, Control, Comparison, Expressing, Cell Culture, Quantitative RT-PCR, Western Blot, Analysis, Morphology