verso complementary dna synthesis kit  (Thermo Fisher)

Journal: bioRxiv

Article Title: Identification of a novel bicarbonate transporter critical for Leishmania virulence

doi: 10.1101/2025.07.16.665125

Figure Lengend Snippet: (A) Expression of LmSLC26A at RNA level. Total RNA was extracted from L. major cell, DNAse treated and cDNA was prepared. The lanes marked as “+RT” and “−RT” represent the reactions with and without RT enzyme, respectively. (B) Topological schematic of LmSLC26A with Transmembrane domain, STAS domain, and Histidine phosphatase domain. The red line represents the region selected for antibody generation. (C) Purification of LmSLC26A 597-935 from bacterial expression system with Ni-NTA affinity purification. WCL- Whole cell lysate, IF- insoluble fraction, SF- soluble fraction, FT- flow through, W- Wash, E- Elute fraction (D) Generation of antibody in mice. Western blot was performed using purified protein with prebleed and test bleed. (E) Generation of antibody in the rabbit. Western blot was performed using purified protein with prebleed and test bleed.

Article Snippet: Total RNA from L. major promastigotes was isolated using Trizol reagent (Invitrogen, Carlsbad, CA, USA), followed by DNase I treatment (Invitrogen). cDNA was prepared from DNase I-treated RNA using the Verso cDNA kit (Thermo Scientific, Waltham, MA, USA) using the manufacturer’s protocol.

Techniques: Expressing, Purification, Affinity Purification, Western Blot