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Plasmids from each sample was purified using commercial plasmid preparation kit (Promega, USA) and sent for sequencing using M13 forward and reverse primers.
Briefly, mRNA from each sample was purified using Sera-mag magnetic oligo dT) beads (Thermo Scientific) [for SUM149PT, Ribominus beads (Invitrogen) was applied to deplete ribosomal RNA species], fragmented at high temperatures into random fragments, and reverse-transcribed into cDNA using Superscript II (Invitrogen).
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RNA of each sample was purified using oligo dT -attached magnetic beads froligo dT -attachedple Prep Kit (Illumagneticn Diego, CA, USA).
The sample was purified using the RNeasy kit.
Genomic DNA from tissue samples was purified using DNAzol (Invitrogen).
DNA from these samples was purified using a commercial Quick Extract Solution kit, following the manufacturer's instructions.
After two hour incubation at 45°C, DNA from the samples was purified using Qiagen miniprep columns (cat. 27106).
PCR products from all positive samples were purified using the PCR Purification Kit (Qiagen, Germantown, USA) and sequenced.
All cDNA probes generated from vascular endothelial biopsy samples were purified using the QIAquick PCR purification kit (Qiagen).
Total RNA from co-cultures and control samples were purified using miRNA Easy Kit (Qiagen, Valencia, CA, USA).
In addition to the RNA samples obtained from oral swabs as described above, RNA samples were purified using Trizol (Invitrogen Corp., Carlsbad, CA).
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