Vd 250r freeze dryer

Ethanol (Wako), acetonitrile (Wako), mEthanol (Wako), sulfuric acid (Wako), acetone (Wako), trifluoroacetate (Tokyo Chemical Industry), dimethyl sulfoxide (Wako), MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide), LPS and Griess reagent were all purchased from Sigma (St. Louis, MO, USA). An ELx 800 Universal Microplate Reader (BIO-TEK), VD-250R Freeze Dryer (TAITEC), US-105 Sonicator (SND), 5420 Centrifuge (IMOTO), R-300 Rotavapor (BUCHI), V-300 Vacuum Pump (BUCHI), High Performance Flash Chromatography (HPFC) system (Biotage AB), Medium Pressure Liquid Chromatography (MPLC) system (EPCLC, Yamazen), Preparative High Performance Liquid Chromatography (PHPLC) system (EPCLC, Yamazen), 1220 Infinity LC (Agilent Technologies), NMR spectrometer (Bruker DRX-600; Bruker Daltonics, Billerica MA, USA), Quadrupole time-of-flight (qTOF) mass spectrometer (Agilent Technologies, USA) and JASCO DIP-370 polarimeter (JASCO, Tokyo, Japan) were used.

The collected dentin shavings were freeze-dried (VD-250R Freeze Dryer, TAITEC, Saitama, Japan) and pulverized at 1500 rpm for 2 min (Shake Master Neo, Bio Medical Science, Tokyo, Japan). The powdered root caries samples were added to a lysis solution (Lysis Solution F, Nippon Gene, Tokyo, Japan) and left to stand for 10 min at 65 °C. The samples were centrifuged at 12,000× g for 1 min, and genomic DNA was extracted using an MPure Bacterial DNA Extraction Kit (MP Biomedical, Santa Ana, CA, USA).

Fecal samples were frozen and dried using VD-250R Freeze Dryer (TAITEC Corp., Saitama, Japan) and were mechanically disrupted using Shake Master Neo (Bristol-Myers Squib. Co., Ltd., NY, USA). Total DNA was extracted using MPure Bacterial DNA Extraction Kit (MP Biomedicals Japan. Co., Ltd., Tokyo, Japan). The concentration of the extracted DNA was determined by using a Synergy H1 (Bio Tek, Instruments, Inc., Winooski, VT, USA) and QuantiFluor dsDNA System (Promega Corp., Madison, WI, USA), and samples were stored at −30°C until use.