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Around 50 mg of dried plant tissue was used for starch quantification, employing a total starch assay procedure kit (Megazyme, Bray, Ireland).
During the procedure Kit with RNase Inhibitor and MultiScribe Reverse Transcriptase was used.
The cDNA library was prepared by using 5 μg of starting material for the Illumina mRNA Sequencing Sample preparation procedure (kit RS-930-1001).
The creatinine analysis was performed by a spectrophotometric method and using a Kinetic Creatinine Procedure Kit provided by Data Medical Associates (Arlington, TX).
Both resistant starch and non-resistant starch were determined according to the enzymatic procedure outlined by the instruction manual for the Megazyme-Resistant Starch Assay Procedure Kit (Megazyme International, Co ., based on the method of McClerry and Monaghan.
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Immunoblots were developed using the enhanced chemiluminescence procedure (ECL kit, Amersham).
DNA was extracted using the 96 well plate procedure (DNeasy 96 kit, Qiagen) according to the manufacturer's directions.
The method of test was in compliance with the procedure of kit.
All 15 participants used the same DNA extraction procedure (QIAamp kit, Qiagen).
mRNA isolation was performed according to standard procedure (Qiagen kit, Venlo, Holland).
For each of the 24 samples, 200 ng of RNA was converted into double-stranded cDNA using the Amino Allyl Message Amp II aRNA amplification procedure (Ambion kit).
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