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). Using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Carlsbad, CA), 1 μg of total RNA was reverse transcribed with random primers. If you have any thoughts regarding the place and how to use manufacturer of sialic acid powder as Raw Material for food, you can make contact with us at the website. Creative Enzymes supplies high quality cholesterol esterase (EC 3.1.1.
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