To confirm if both α2,3 and α2,6 sialic acids facilitate AAV1 and AAV6 transduction, we carried out a lectin competition assay on these three cell lines (Fig. 5A to C). In competitive binding assays, 10 mm free synthetic sialic acid (Sigma) was added 30 min before the endocytic agent and left during the assay. The supernatants were removed and used for total sialic acid quantification. Total RNA was extracted using the RNeasy Mini Kit and the RNase-Free DNase Set to eliminate genomic DNA, all from Qiagen (Manchester, UK). If you have any sort of inquiries pertaining to where and how you can make use of manufacturer of sialic acid powder for food Ingredients, you could contact us at the web-page. Using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Carlsbad, CA), 1 μg of total RNA was reverse transcribed with random primers.
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