Generuler 50 base pair dna ladder

Poly(A) Tail-Length Assay Kit (Thermo Fisher, 764551KT) was used according to the manufacturer’s instructions. PCR products corresponding to the tail- and gene-specific primer combinations of mouse 16S rRNA (universal forward primer: 5′-GGTCGGTTTCTATCTATTTACGATTTCTC-3′, gene-specific reverse primer: 5′-TTCTCTAGGTTAGAGGGTGTACGTATAT-3′) and human ACTB (assay control, primer composition not disclosed by manufacturer) were loaded on a 2.5% agarose gel (Lonza, 50010) and stained with GelRed (Merck, SCT123). Product sizes determined using the GeneRuler 50-base-pair DNA ladder (Thermo Scientific, SM0371). Subsequently, tail- and gene-specific PCR products of mouse 16S rRNA were purified by gel-elution (Cytiva Life Sciences, 28903470) and sent for Sanger sequencing with the shared forward primer (5′-GGTCGGTTTCTATCTATTTACGATTTCTC-3′). Resulting chromatograms were analyzed using SnapGene (v.6.0.2). After confirming alignment to the reference sequence, the unclipped chromatograms were used to visualize 3′ ends.