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Amount of total phenolic contents were assessed using Folin-Ciocalteu reagent procedure [32].
The extract and fractions of stem, root and leaves exhibited TPC values as was measured by Folin-Ciocalteu reagent procedure in the range of 50.80-735.11 50.80-735.111, 55.02-705.11 (Gallic acid equivalent, (GAE) mg/100 g) of dry extract, respectively.
Total RNA was extracted using the TRIzol Reagent procedure (Invitrogen).
Cells were transfected according to the jet prime reagent procedure (Poly plus transfection, Pontoise, France).
The transfection of siRNAs was performed using the Oligofectamine™ reagent procedure.
Presence of sugar in each fraction was determined colorimetrically by the anthrone reagent procedure.
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Tissue RNA was extracted using the tri-reagent procedure according to the manufacturer protocol (Molecular Research Center).
Total RNA and DNA were extracted from 2 × 10 cells by using Tri-reagent procedure (Sigma Chem. Co). and re-suspended in 50 μl of DEPC water.
The limitations of this assay are largely related to the difficulty in interpreting the literature due to lack of standardisation of assay reagents, procedures and scoring.
Prosigna used the 212 UNC and WashU training samples to generate a static control as all four subtypes were well represented in this population with gene expression values generated on the same platform using the same clinical grade reagents, procedures, and the same normalization scheme.
The CFT test proper and reagent preparation procedures followed the procedures outlined by the OIE [ 21].
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