Abstract Purpose To validate a simplified RNA isolation method from biofabricating hydroxyapatite (HAp) scaffolds seeded with mesenchymal stem cells (MSCs) and to identify the appropriate reference gene. Methods Ten MSCs-HAp composites were used for by methods based on homogenization steps column-based purification procedures, while remaining (n = 13) was extracted traditional single-step metho...

Reverse transcription and real-time PCR have become the method of choice for the detection of low-abundance mRNA transcripts obtained from small human muscle biopsy samples. GAPDH, beta-actin, beta-2M, and 18S rRNA are widely employed as endogenous control genes, with the assumption that their expression is unregulated and constant for given experimental conditions. The aim of this study was to...

lentiviral vectors (lvs) are useful vehicle for genetransfer to dividing and non-dividing cells and genetic manipulations. however, the use of lentiviruses in studies requires an accurate titration technique.quantitative real-time pcr (qpcr) is a sensitive technique for the indication and quantitation of retrovirals particles. in this study, we used the qpcr for lentiviral vector titeration. th...

Fusarium wilt (caused by oxysporum f. sp. Lilii) is one of the most damaging diseases in lily (Lilium sargentiae Wilson). Although some F. oxysporum-resistant varieties have been identified and are being utilized resistant breeding, regulation network resistance-associated mechanisms yet to be studied due lack reliable reference genes for qRT-PCR (quantitative reverse transcription PCR) normali...

Quantification by real-time RT-PCR requires a stable internal reference known as a housekeeping gene (HKG) for normalising the mRNA levels of target genes. The present study identified and validated stably expressed HKGs in post-thaw Symbiodinium clade G. Six potential HKGs, namely, pcna, gapdh, 18S rRNA, hsp90, rbcl, and ps1, were analysed using three different algorithms, namely, GeNorm, Norm...

Members of the tumor necrosis factor (TNF) superfamily have been revealed to be associated with painful bladder syndrome/interstitial cystitis (PBS/IC). TNF ligand-related molecule 1A (TL1A) and its receptor, death receptor 3 (DR3), belong to the TNF superfamily and have been implicated in chronic inflammatory diseases. Bladder biopsies from 8 female patients clinically diagnosed with PBS/IC ac...

The choice of reference genes that are stably expressed amongst treatment groups is a crucial step in real-time quantitative PCR gene expression studies. Recent guidelines have specified that a minimum of two validated reference genes should be used for normalisation. However, a quantitative review of the literature showed that the average number of reference genes used across all studies was 1...

Abstract Purpose Herpes simplex diffuse endotheliitis with accompanying feathery infiltration is difficult to diagnose due corneal findings that are similar fungal keratitis. This case series reports on the effectiveness of using real-time polymerase chain reaction (PCR) herpes in appearance Methods After extracting smear sample DNA, samples were then applied two independent PCR assays, a quali...