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Formatting, expression and purification of nanobodies with ncaa incorporated The insertion of ncAA is achieved by mutating the sequence to a stop codon UAG at a specific site in the protein

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This can happen from normal aging (we lose 10-20% of collagen per decade after 30) or by over-stretching the collagen layer and creating rips in it

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For the BSF control, 1 mL of 50% BSF/PBS without inoculation was transferred to a 50 mL conical tube and similarly 1:1:1 methanol / ddH 2 O / chloroform (Fisher) was added for a total volume of 24 mL