Gel doctm ez

The binding capacity of TROPY toward DNA was analyzed in reaction mixtures containing 100 ng of FAM-HpScr9 and increasing amounts of TROPY in a final volume of 10 µL. After 20 min at room temperature, the samples were electrophoresed in 0.8% agarose gels in TAE buffer 1X. Gels were visualized on Gel Doc^TM EZ (Bio-Rad Laboratories, Inc., Barcelona, Spain).

Total nucleic acid from the MDR K. pneumoniae isolates was extracted through WizPrepTM gDNA Mini Kit (Seongnam-si, Gyeonggi-do, 13209, Republic of Korea). Molecular characterization of four (04) capsular encoding genes; serotype K1, serotype K2, serotype K5, and serotype K54 was performed through PCR by using gene-specific primers, as given in Table 2. PCR Primer-specific fragments were visualized in 01% agarose gel electrophoresis using Gel Doc^TMEZ (BIO-RAD).

Molecular identification of CK1-CK5 was carried out using multiplex PCR. The nucleotide sequences of the selected region of the authentic medicinal plants in this study together with other medicinal plants in the genera Betula, Strychnos, and Ziziphus from the GenBank database were aligned (S1 Spreadsheet) to search the different sites in order to design species-specific primers. Three sets of species-specific primers (Table 2), i.e., KSKITSF/BeAITS2R, KSKITSF/StAITS2R, and KSKITSF/ZiAITS2R, as well as an internal control primer, KSKITSF/ITS4, were utilized simultaneously in multiplex PCR amplification. Strychnos nux-blanda A.W. Hill (Strychnaceae), which was authenticated by the botanist and kept in the herbarium (voucher specimen no. YS19-StN1), was used as a negative control. The reaction mixture and PCR amplification conditions used were the same as those used in the aforementioned protocol. The amplicons were finally determined by 2.2% agarose gel electrophoresis and captured on a UV tray using an imager (Gel Doc^TM EZ, Bio-Rad Laboratories, USA) and Image Lab version 3.0 software.