Quantifying Pneumocystis Fungal Burden

The right lung was placed in 1 mL of Trizol and briefly stored at −20°C. RNA was extracted from each lung following the manufacturer’s protocol. RNA was quantified using a Nanodrop spectrophotometer. cDNA was synthesized from 1 μg of Lung RNA using BioRad Reverse Transcription Supermix for RT-qPCR following the manufacturer’s protocol. Pneumocystis fungal burden was assayed via amplifying a 166-bp fragment of the small ribosomal rRNA (ssRNA) from the mitochondrial genome, as described previously (6 (link), 7 (link)).

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de la Rua N.M., Samuelson D.R., Charles T.P., Welsh D.A, & Shellito J.E. (2016). CD4+ T-Cell-Independent Secondary Immune Responses to Pneumocystis Pneumonia. Frontiers in Immunology, 7, 178.

Publication 2016

Reverse Transcription Supermix

Cdna Lung Mitochondrial genome Pneumocystis Reverse transcription Ribosomal Rrna Trizol

Corresponding Organization :

Other organizations : Louisiana State University Health Sciences Center New Orleans

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Pneumocystis fungal burden

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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