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All related inserts were available in the Gateway® entry-vector (pENTR1A) for recombination into the destination vectors by using the "LR-clonase" enzyme kit (Invitrogen) [38].
All the further experimental steps including the transformation of E.coli DH5alpha were done according to the instructions of the Gateway Clonase LR II enzyme kit (Invitrogen).
First-strand cDNA was synthesized from 1 µg total RNA using the Superscript II enzyme kit (Invitrogen) and random hexamer primers.
The concentration of amylase was measured by an amylase activity enzyme kit (Abcam, Cambridge, UK).
A commercial enzyme kit (Invitrogen Superscript III) was used to synthesize cDNA.
Alkaline phosphatase was measured by enzymatic colorimetry using a Pars Azmoon kit (Pars Azmoon, Iran); Serum calcium and phosphorous were both measured by colorimetery using the Kavoshyar enzyme kit (Kavoshyar, Iran) and Sheem enzyme kit (Sheem enzyme, Iran), respectively.
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Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) enzyme kits were obtained from Bio-Rad (UK).
Cholesterol, TAG, HDL cholesterol and LDL cholesterol in serum were analysed using colorimetric enzyme kits (Roche Diagnostics).
Total cholesterol TC in mg/dL, HDLc (mg/dL), and triglycerides (mg/dL) were determined using colorimetric enzyme kits (Spinreact, Bas Gerona, Spain).
All biological reagents, including enzymes, kits, and biomaterials, are listed in Additional file 3: Table S1.
CRP concentrations were determined by enzyme immunoassay kit (Virgo CRP150 kit, Hemagen, Waltham, MA, USA).
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