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Publications (10)
We recently have shown that the gut microbiota composition in female and male runners positively correlates with sports, and female runners show similar gut microbiome diversity to male runners. However, gut microbiota composition has not yet been fully investigated in other endurance athletes, such as cyclists. Therefore, in the current study, we...
There is still a pressing need for further investigation to bridge the gap in understanding the differences in gut microbiota composition between female runners and their male counterparts. We aimed to determine the gut microbiota composition in competitive non-professional female and male runners and to correlate the gut bacteria to performance. O...
Objective
Single‐nucleotide variants (SNVs) are of great significance in prenatal diagnosis as they are the leading cause of inherited single‐gene disorders (SGDs). Identifying SNVs in a non‐invasive prenatal screening (NIPS) scenario is particularly challenging for maternally inherited SNVs. We present an improved method to predict inherited SNVs...
Background: Immediate early genes (IEGs) are rapidly activated and initiate diverse cellular processes including neuroplasticity. We report the effect of psilocybin (PSIL), PSIL-containing psychedelic mushroom extract (PME) and 5-hydroxytryptophan (5-HTP) on expression of the IEGs, cfos, egr1, and egr2 in mouse somatosensory cortex (SSC).
Methods:...
Delta-9-tetrahydrocannabinol (THC) is the primary psychoactive compound of the cannabis plant and an exogenous ligand of the endocannabinoid system. In previous studies, we demonstrated that a single microdose of THC (0.002 mg/kg, 3–4 orders of magnitude lower than the standard dose for rodents) exerts distinct, long-term neuroprotection in model m...
Williams syndrome (WS) is a multisystem neurodevelopmental disorder caused by a de novo hemizygous deletion of ~26 genes from chromosome 7q11.23, among them the general transcription factor II-I (GTF2I). By studying a novel murine model for the hypersociability phenotype associated with WS, we previously revealed surprising aberrations in myelinati...
Ricin, derived from the castor bean plant, is a highly potent toxin, classified as a potential bioterror agent. Current methods for early detection of ricin poisoning are limited in selectivity. MicroRNAs (miRNAs), which are naturally occurring, negative gene expression regulators, are known for their tissue specific pattern of expression and their...
Questions
Questions (2)
Hi,
It is not the first time I've used TaqMan miRNA assays to quantitate the relative change of a miR of interest (usually with U6 as calibrator), and it has always worked fine. However, in my last experiment, my miR Cts were extraordinarily high- between 33 and 38+, some even undetectable.
I have used the same protocol as I've always had, and have a feeling that these results are due to low expression of this miR in the investigated tissue (a feature that cannot be changed).
For the RT part, I use reagents from the ThermoFisher HighCapacity kit, with TaqMan RT primers. In this reaction, I put 5ul of 2ng/ul total RNA, which, according to the company's protocol, is the maximal amount of RNA per this reaction (15ul in total, including primers and reagents).
In the real-time qPCR part, I put 2ul of the RT product in a 20ul reaction well. This resulted in the unusually high Cqs. I should mention that I've never investigated this miR in this tissue, so it is possible that its expression is low there. To overcome the problem, I tried to put 4ul of the RT product (instead of 2ul as before) in the 20ul qPCR reaction, but the Cts did not change dramatically. Unfortunately, I have reached the cDNA limit according to the TaqMan protocol.
Has anyone ever tried using a greater amount of RNA in the RT reaction, disregarding the official protocol?
Thanks in advance
Hi,
I am planning to overexpress a specific miR (it's 5p strand)in cortical neurons of mice (in vivo) using AAV2 infection. In order for the process to be more natural, I wanted to use the pre-miR, so it will undergo processing in the cell.
The problem is that there are two prevalent precursors: the more prevalent expresses both 3p and 5p, while the less prevalent (but still common) expresses the 5p alone.
Which one should I use? Or maybe I should consider forgoing the pre-miR protocol and sticking with the mature miR alone?
Thanks.
