Taqman fast advanced reagents

Manufactured by Thermo Fisher Scientific

TaqMan™ Fast Advanced reagents are a collection of real-time PCR reagents designed for rapid and sensitive detection of target DNA sequences. The reagents provide efficient amplification and detection, enabling fast and reliable quantitative analysis.

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Lab products found in correlation

Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) Maxwell rsc simply rna blood kit, by Promega (1 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Viia 7 real time pcr system, by Thermo Fisher Scientific (1 mentions) Maxwell rsc instrument, by Promega (1 mentions) Magna pure lc, by Roche (1 mentions)

Close spelling variants of this product

TaqMan reagents (162 citations) Taqman fast reagents (5 citations)

3 protocols using taqman fast advanced reagents

Quantitative Gene Expression Analysis

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Total RNA was extracted from two Transwell inserts per condition using the Maxwell® RSC simplyRNA Blood kit on the Maxwell® RSC instrument (both Promega) and transcribed into cDNA using the High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). Expression of genes of interest relative to the expression of two housekeeping genes (ATP5B, YWHAZ) was determined using TaqMan™ Fast Advanced reagents on a ViiA 7 Real-Time PCR system (both Applied Biosystems). TaqMan™ assay IDs are listed in Table 2.Table 2

TaqMan™ assays used for gene expression analysis.

Gene	Assay ID	Gene (cont.)	Assay ID (cont.)
ACE2 (full-length isoform)	Hs00222343_m1	IFIT1	Hs00356631_g1
AREG	Hs00950669_m1	IFNB1	Hs01077958_s1
ATP5B (housekeeping)	Hs00969569_m1	IFNL1	Hs00601677_g1
BTC	Hs01101204_m1	IL6	Hs00174131_m1
CTSL	Hs00964650_m1	ISG15	Hs01921425_s1
CXCL10	Hs01124252_g1	KRT5	Hs00361185_m1
CXCL8	Hs00174103_m1	MUC5AC	Hs01365616_m1
DDX58	Hs01061436_m1	MUC5B	Hs00861595_m1
EGF	Hs01099990_m1	NRP1	Hs00826128_m1
EGFR	Hs01076090_m1	NRP2	Hs00187290_m1
EPGN	Hs02385424_m1	TGFA	Hs00608187_m1
EREG	Hs00914313_m1	TJP1	Hs01551861_m1
FOXJ1	Hs00230964_m1	TMPRSS2	Hs01122322_m1
FURIN	Hs00965485_g1	TNF	Hs00174128_m1
HBEGF	Hs00181813_m1	YWHAZ (housekeeping)	Hs01122445_g1
IFIH1	Hs00223420_m1

Stölting H., Baillon L., Frise R., Bonner K., Hewitt R.J., Molyneaux P.L., Gore M.L., Barclay W.S., Saglani S, & Lloyd C.M. (2022). Distinct airway epithelial immune responses after infection with SARS-CoV-2 compared to H1N1. Mucosal Immunology, 15(5), 952-963.

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Chlamydiae and Ichthyocystis qPCR Protocols

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A modified Chlamydiae-specific Taqman qPCR protocol (Lienard et al., 2011 (link)) (forward: 5′-CCGCCAACACTGGGACT-3′, reverse: 5′-GGAGTTAGCCGGTGCTTCTTTAC-3′, probe: 5′-FAM-CTACGGGAGGCTGCAGTCGAGAATC-BHQ-3′) was used. A qPCR protocol designed for the Ca. Ichthyocystis genus (forward: 5′-CAAGGCGACGATCGGTAGCTG-3′, reverse: 5′-TTACAACCCTAAGGCCTTCTTCACC-3′, probe: 5′-FAM-TTGCTGGATCAGGCTTCCGCCCATTGTCCAAA-BHQ-3′) allowed quantification of high concentrations of bacteria within infected material. All qPCR protocols were validated against serial dilutions of amplicons from target and control species. Reactions were carried out on a StepOne Plus real-time PCR system (Applied Biosystems) using Taqman Fast Advanced reagents (Applied Biosystems) according to the manufacturer’s instructions with 1-5 μl DNA in a reaction volume of 20 μl.

Seth-Smith H.M., Dourala N., Fehr A., Qi W., Katharios P., Ruetten M., Mateos J.M., Nufer L., Weilenmann R., Ziegler U., Thomson N.R., Schlapbach R, & Vaughan L. (2016). Emerging pathogens of gilthead seabream: characterisation and genomic analysis of novel intracellular β-proteobacteria. The ISME Journal, 10(7), 1791-1803.

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Quantifying W. chondrophila Infection in Larvae

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A qPCR system was designed against the 16S rRNA gene from W. chondrophila (forward: 5′- AGTCCGGCTACACCAAGTATGC-3′, reverse: 5′-TGGCGAAGGCGGTTTTC-3′, probe: 5′-FAM-TTCGCTCCCCTAGCTTTCGGGCAT-TAMRA-3′), allowing quantification of the bacterial load of individual infected larvae. The TaqMan qPCR system was designed and validated against quantified serial dilutions of the target sequence cloned into the pCR2.1 plasmid and against total DNA extracts of non-infected larvae. Serial dilutions of pCR2.1 containing the target sequence were used as standards in each run. Total DNA of individual larvae was extracted with a MagNA Pure LC (Roche) robot and eluted in 100 μl elution buffer. Reactions were carried out on a StepOne Plus real-time PCR system (Applied Biosystems). TaqMan Fast Advanced reagents (Applied Biosystems) were used according to the manufacturer's instructions with 5 μl input DNA in a total reaction volume of 20 μl.

Fehr A.G., Ruetten M., Seth-Smith H.M., Nufer L., Voegtlin A., Lehner A., Greub G., Crosier P.S., Neuhauss S.C, & Vaughan L. (2016). A Zebrafish Model for Chlamydia Infection with the Obligate Intracellular Pathogen Waddlia chondrophila. Frontiers in Microbiology, 7, 1829.

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