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PCR products of analyzed genes were sequenced to validate the correct analysis of gene targets.
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Based on this analysis, a total of 289 gene products (36% of analyzed mRNA-protein pairs) displayed a difference of two or more between the values of PC-1 in mRNA and protein domains (|PC-1mRNA−PC-1protein|≥2).
Following DNA amplification, PCR products of markers analyzed by sequencing were purified using the QIAquick PCR purification kit (Qiagen, Hilden, Germany).
Four of the patients (No. 2 to No. 5) had their products of conception analyzed while patient No. 1 had no cytogenetic analysis available due to an early spontaneous abortion.
The small number of products analyzed cannot be representative nor comprehensive for natural cosmetics in general.
The quantification of products was analyzed using the ImageQuant TL software (GE Healthcare).
Chemical composition of products was analyzed using a gas chromatography (Agilent GC-6890) with a HP-FFAP silica capillary column (30 m × 0.32 mm × 0.25 μm).
Structures of products were analyzed by X-ray diffractometer (XRD, Shimadzu XRD 6000, Nakagyo-ku, Kyoto, Japan) and transmission electron microscope (TEM, JEOL-2010, JEOL Ltd., Akishima, Tokyo, Japan).
The DNA topology of products was analyzed on 1% agarose gels in 40 mM Tris-acetate (pH 8.0) and 1 mM EDTA containing 1 μg/mL ethidium bromide.
Following amplification, melting temperatures of PCR products were analyzed to determine specificity of the PCR product.
This approach has particular advantages over other methods of analyzing products of microbial cell culture.
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