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    Millipore rnalater
    Preliminary GLST (multiplex) trials on T. cruzi I mock infections. We created mock infections by mixing 10 4 , 10 5 and 10 6 <t>RNAlater-preserved</t> TcI-Sylvio epimastigote (epi) cells with uninfected Rhodnius prolixus vector gut (UVG). DNA extracted from these mock infections was subjected to the multiplexed, 203-target GLST reaction (using the same cycling conditions as for single-target reactions – see Methods or Supplementary Fig. 2 legend) and products were electrophoresed in 0.8% agarose gel. Fainter banding of GLST products from lower concentration mock infections encouraged follow-up on sensitivity thresholds using additional dilution curves and qPCR. Next to DNA ladder (L) and no-template control (NTC), the gel also contains TcZ primer product from pure TcI epimastigote DNA. TcZ primers provide a highly sensitive positive control (PC) as they target 195 bp satellite DNA repeats that make up ca. 5% of the T. cruzi genome.
    Rnalater, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 3603 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Preliminary GLST (multiplex) trials on T. cruzi I mock infections. We created mock infections by mixing 10 4 , 10 5 and 10 6 RNAlater-preserved TcI-Sylvio epimastigote (epi) cells with uninfected Rhodnius prolixus vector gut (UVG). DNA extracted from these mock infections was subjected to the multiplexed, 203-target GLST reaction (using the same cycling conditions as for single-target reactions – see Methods or Supplementary Fig. 2 legend) and products were electrophoresed in 0.8% agarose gel. Fainter banding of GLST products from lower concentration mock infections encouraged follow-up on sensitivity thresholds using additional dilution curves and qPCR. Next to DNA ladder (L) and no-template control (NTC), the gel also contains TcZ primer product from pure TcI epimastigote DNA. TcZ primers provide a highly sensitive positive control (PC) as they target 195 bp satellite DNA repeats that make up ca. 5% of the T. cruzi genome.

    Journal: bioRxiv

    Article Title: Genome-wide locus sequence typing (GLST) of eukaryotic pathogens

    doi: 10.1101/2020.03.24.003590

    Figure Lengend Snippet: Preliminary GLST (multiplex) trials on T. cruzi I mock infections. We created mock infections by mixing 10 4 , 10 5 and 10 6 RNAlater-preserved TcI-Sylvio epimastigote (epi) cells with uninfected Rhodnius prolixus vector gut (UVG). DNA extracted from these mock infections was subjected to the multiplexed, 203-target GLST reaction (using the same cycling conditions as for single-target reactions – see Methods or Supplementary Fig. 2 legend) and products were electrophoresed in 0.8% agarose gel. Fainter banding of GLST products from lower concentration mock infections encouraged follow-up on sensitivity thresholds using additional dilution curves and qPCR. Next to DNA ladder (L) and no-template control (NTC), the gel also contains TcZ primer product from pure TcI epimastigote DNA. TcZ primers provide a highly sensitive positive control (PC) as they target 195 bp satellite DNA repeats that make up ca. 5% of the T. cruzi genome.

    Article Snippet: Insects were euthanized with CO2 and hindguts drawn into 5 volumes of RNAlater (Sigma-Aldrich) by pulling the abdominal apex toward the posterior with sterile watchmaker’s forceps.

    Techniques: Multiplex Assay, Plasmid Preparation, Agarose Gel Electrophoresis, Concentration Assay, Real-time Polymerase Chain Reaction, Positive Control

    Comparison of extracted RNA of RNAlater immersed and formalin-fixed paraffin-embedded (FFPE) oviductal cell populations using the Experion system. The extracted RNA of RNAlater immersed (A–C) and FFPE (D–F) oviductal cell populations were

    Journal: Journal of Histochemistry and Cytochemistry

    Article Title: Cellular Expression and Localization of Estrogen Receptor a and Progesterone Receptor mRNA in the Bovine Oviduct Combining Laser-Assisted Microdissection, Quantitative PCR, and In Situ Hybridization

    doi: 10.1369/0022155410397995

    Figure Lengend Snippet: Comparison of extracted RNA of RNAlater immersed and formalin-fixed paraffin-embedded (FFPE) oviductal cell populations using the Experion system. The extracted RNA of RNAlater immersed (A–C) and FFPE (D–F) oviductal cell populations were

    Article Snippet: RNA isolation from RNAlater immersed tissue was performed using the Tri Reagent isolation kit (Sigma-Aldrich, Munich, Germany) according to the manufacturer’s protocol.

    Techniques: Formalin-fixed Paraffin-Embedded