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RNA-guided targeting of genes in human cells was achieved through coexpression of the Cas9 protein with gRNAs using reagents prepared by the Church group (Mali et al., 2013), which are available from the Addgene (http://www.addgene.org/crispr/church/).org/crispr/church/
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The beads used to determine cytokine levels were prepared by using reagents described in Table 1.
Samples were prepared by normalizing each to equivalent volumes using phosphate buffered saline followed by purification using reagents to extract impurities and isolate pathogenic GAG (pGAG).
All the reagents and solutions were prepared using reagent-grade chemicals from Sigma Aldrich (South Africa) without further purification.
Temperature-sensitive ceramics were prepared by a conventional ceramic processing route using reagent grade cooper carbonate hydroxide and nickel (cobalt) carbonate hydroxide hydrates [11].
A blank reagent was prepared by using 1 mL of water and 4 mL of anthrone reagent in a test tube and treated similarly.
The Dragendorf reagent was prepared by mixing two reagents: reagent 1 and reagent 2 in equal parts.
The reagents were prepared by polymer analogous conversions using polystyrene crosslinked with ethyleneglycol dimethacrylate as the carrier matrix.
Dye reagent was prepared by diluting 1 part dye reagent (Bio-Rad) concentrate with 4 parts Milli-Q water.
Copper reagent was prepared by preparing an aqueous solution of copper (II) acetate monohydrate (5% w/v) and adjusting the pH to 6.1 by using pyridine.
All the reagents, like buffers and loading dye, used in RAPD were prepared by the standard method of Nayak et al. (2003).
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