Pre designed primer probe mixes

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

Pre-designed primer/probe mixes are a type of molecular biology laboratory equipment used for the amplification and detection of specific DNA or RNA sequences. They contain pre-optimized oligonucleotide sequences necessary for performing PCR or RT-PCR experiments.

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Lab products found in correlation

Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions) Random hexamer, by Thermo Fisher Scientific (1 mentions) Steponeplus real time pcr machine, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

Pre-designed primers Pre-designed probes Primer-probe mixes 20X primer-probe mix Designed primers Primer Mix Primer probes

2 protocols using pre designed primer probe mixes

Quantitative Gene Expression in Intervertebral Disc

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Target genes were investigated using quantitative real-time polymerase chain reaction (qRT-PCR) conducted on 10 non-degenerate AC samples (microscopic grade 0-6), 11 non-degenerate NP samples (grade <4), 24 moderately degenerate NP samples (grade 4-7), 18 severely degenerate NP samples (grade >7) and 18 NP samples with evident infiltration (Supplementary Table I, II, III).
qRT-PCR was performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems, Lutterworth UK) for potential NP markers (Pre-designed primer/probe mixes Applied Biosystems) (Supplementary Table IV). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 18S (Applied Biosystems) were used as housekeeping genes. Ten microliter reactions were prepared using TaqMan Universal PCR Master Mix (Applied Biosystems). Results were analysed using the 2^−ΔCt method and presented as relative gene expression normalised to the average C_T for the two housekeeping genes.

Thorpe A.A., Binch A.L., Creemers L.B., Sammon C, & Le Maitre C.L. (2015). Nucleus pulposus phenotypic markers to determine stem cell differentiation: fact or fiction?. Oncotarget, 7(3), 2189-2200.

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Quantifying Gene Expression in NP Cells

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Following stimulation NP cells were recovered from alginate culture and re-suspended in 0.06%w/v type I collagenase and incubated for 10 minutes at 37°C and cells recovered by centrifugation. RNA was extracted using Qiagen RNeasy Mini kit (Qiagen, Crawley, UK) as per manufacturers' protocol. cDNA was reverse transcribed using Moloney Murine Leukaemia Virus reverse transcriptase (Bioline) and random hexamers (Applied Biosystems, Warrington, UK). cDNA samples were interrogated by qRT-PCR for the expression of MMP3, MMP13, IL-1β, IL-6, IL-8 and aggrecan (Pre-designed primer/probe mixes Applied Biosystems). Plates were ran on a StepOnePlus real-time PCR machine for 50 cycles (Applied Biosystems). Relative gene expression levels were calculated using the 2 -ΔΔCT method and normalised against two internal reference genes (GAPDH and 18S) and unstimulated control samples. Stable expression of internal reference genes was confirmed by geNorm algorithm.

Daniels J., Binch A.A, & Le Maitre C.L. (2017). Inhibiting IL-1 signaling pathways to inhibit catabolic processes in disc degeneration. Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 35(1).

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