Relative Expression Levels of adeB, ompA, and ompW in A. baumannii

The relative expression levels of adeB, ompA, and ompW from A. baumannii were evaluated using 1 µg of purified total RNA as a template for reverse transcription, which was performed using the High-Capacity cDNA Reverse Transcription kit (Invitrogen). Amplification of the target genes was carried out using previously described oligonucleotide primers [46 (link), 47 (link)]. To normalize the gene expression levels, the single copy housekeeping gene rpoB was used [47 (link)]. The qPCR analysis was performed in duplicate using the Platinum® SYBR® Green qPCR SuperMix-UDG kit (Invitrogen) in a StepOne thermocycler (Applied Biosystems, Massachusetts, USA).

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Schmitt B.L., Leal B.F., Leyser M., de Barros M.P., Trentin D.S., Ferreira C.A, & de Oliveira S.D. (2023). Increased ompW and ompA expression and higher virulence of Acinetobacter baumannii persister cells. BMC Microbiology, 23, 157.

Publication 2023

High-Capacity cDNA Reverse Transcription kit Platinum® SYBR® Green qPCR SuperMix-UDG kit StepOne thermocycler

Cdna Gene expression Genes amplification Housekeeping gene Oligonucleotide primers Platinum Reverse transcription Sybr green

Corresponding Organization :

Other organizations : Pontifícia Universidade Católica do Rio Grande do Sul, Universidade Federal de Ciências da Saúde de Porto Alegre

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Relative expression levels of adeB, ompA, and ompW from A. baumannii

control variables

1 µg of purified total RNA as a template for reverse transcription
The High-Capacity cDNA Reverse Transcription kit (Invitrogen) used for reverse transcription
Previously described oligonucleotide primers [46, 47] used for amplification of target genes
The single copy housekeeping gene rpoB used to normalize the gene expression levels [47]
The qPCR analysis performed in duplicate using the Platinum® SYBR® Green qPCR SuperMix-UDG kit (Invitrogen) in a StepOne thermocycler (Applied Biosystems, Massachusetts, USA)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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