aurum high stringency wash  (Bio-Rad)

Journal: PLoS ONE

Article Title: Molecular and Cellular Characterization of an AT-Hook Protein from Leishmania

doi: 10.1371/journal.pone.0021412

Figure Lengend Snippet: (A) Schematic of the entire open reading frame predicted by LmjF06.0720. GR indicates Glycine-Arginine dipeptide repeats; ATH denotes the classical RGRP AT-hook motif; YEATS, indicates YEATS protein family domain. (B) Sequence alignment of the first ∼330 amino-acids in LmjF06.0720 and its orthologs in L. amazonensis , L. mexicana, L. infantum and L. braziliensis . GR1, GR2 and ATH are emphasized in red (C) Semi-quantitative analysis of LamAT-Y expression. Total RNA was isolated from L. amazonensis promastigotes and axenic amastigotes and quantified prior to cDNA synthesis. The cDNAs were then amplified by PCR for 30 cycles with input amounts of 3, 8 and 25 ng RNA as denoted using LamAT-Y-specific and P4 nuclease-specific primers and analysed by agarose gel electrophoresis as indicated. Lane 25* indicates PCRs performed with RNA templates that were not reverse-transcribed.

Article Snippet: For quantitative real-time PCR, RNA was extracted from parasites using the Aurum total RNA extraction system (BioRad), following which 5 µM of RNA was reverse-transcribed using the iScript cDNA synthesis kit (BioRad).

Techniques: Sequencing, Expressing, Isolation, Amplification, Agarose Gel Electrophoresis