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RNA extraction and miRNA detection For the isolation and purification of total RNA, including small RNAs from SHSY5Y cells, in the first step, trypsin was used to detach the cells from the flask and collected in 1.5 ml tubes

42 While a severe loss of activity (10000-fold) was reported when it was substituted for Ala, 29 replacement with Glu led to only a moderate loss in affinity (31-fold), confirming the requirement of a negatively charged amino acid at position 15

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