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Taqman preamp master mix protocol

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The TaqMan PreAmp Master Mix protocol is a laboratory reagent designed to preamplify target nucleic acid sequences prior to real-time PCR analysis. It enables the amplification of low-copy number targets by increasing the starting template quantity, thereby improving the sensitivity and detection of qPCR assays.

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Lab products found in correlation

Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Data assist 3, by Thermo Fisher Scientific (2 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (2 mentions) Sds 2, by Thermo Fisher Scientific (2 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions) Taqman reverse transcription reagent, by Thermo Fisher Scientific (1 mentions) Rnalater, by Merck Group (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions)

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TaqMan Master Mix (448 citations) TaqMan PreAmp Master Mix (315 citations) PreAmp Master Mix (17 citations) TaqMan® PreAmp (8 citations) TaqMan® PreAmp Master Mix Kit protocol (5 citations)

3 protocols using taqman preamp master mix protocol

1

Quantification of TGF-β1 Expression

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Primary pup AM (200,000 cells) were stored in RNALater (Sigma, St. Louis, MO) at −80°C. mRNA was purified using a commercially available RNA isolation kit (RNEasy Plus Mini Kit, Qiagen, Valencia, CA) according to the manufacturer's instructions. cDNA was synthesized with oligo‐dT primers using TaqMan Reverse Transcription Reagents (Applied Biosystems, Foster City, CA) according to the manufacturer's protocol. TGFβ1 was preamplified using TaqMan PreAmp Master Mix Protocol (Applied Biosystems, Foster City, CA). Following a 1:20 dilution of the preamplified cDNA product, TaqMan real‐time quantitative PCR was performed using the TaqMan Universal Master Mix protocol and the TGFβ1 TaqMan assay (Applied Biosystems) on a StepOne Plus Real‐Time PCR system. Data were analyzed using the ΔΔCT method with GUSB and B2M as housekeeping genes.
Gauthier T.W., Grunwell J.R., Ping X., Harris F.L, & Brown L.A. (2017). Impaired defenses of neonatal mouse alveolar macrophage with cftr deletion are modulated by glutathione and TGFβ1. Physiological Reports, 5(6), e13086.
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2

Quantifying Gene Expression in Hypothalamus

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For gene-expression analyses, mRNA from microdissected hypothalamic explants or FACS-sorted tanycytes was reverse transcribed using SuperScript III Reverse Transcriptase (Life Technologies) and a linear preamplification step applied to sorted cells only using the TaqMan PreAmp Master Mix protocol (P/N 4366128, Applied Biosystems). RT-PCR was carried out on the Applied Biosystems 7900HT Fast Real-Time PCR System using exon-boundary-specific TaqMan Gene Expression Assays (Applied Biosystems) (Table S1). Gene-expression data were analyzed using SDS 2.4.1 and Data Assist 3.0.1 software (Applied Biosystems).
Duquenne M., Folgueira C., Bourouh C., Millet M., Silva A., Clasadonte J., Imbernon M., Fernandois D., Martinez-Corral I., Kusumakshi S., Caron E., Rasika S., Deliglia E., Jouy N., Oishi A., Mazzone M., Trinquet E., Tavernier J., Kim Y.B., Ory S., Jockers R., Schwaninger M., Boehm U., Nogueiras R., Annicotte J.S., Gasman S., Dam J, & Prévot V. (2021). Leptin brain entry via a tanycytic LepR:EGFR shuttle controls lipid metabolism and pancreas function. Nature metabolism, 3(8), 1071-1090.
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3

Quantifying Gene Expression in Hypothalamus

Check if the same lab product or an alternative is used in the 5 most similar protocols
For gene-expression analyses, mRNA from microdissected hypothalamic explants or FACS-sorted tanycytes was reverse transcribed using SuperScript III Reverse Transcriptase (Life Technologies) and a linear preamplification step applied to sorted cells only using the TaqMan PreAmp Master Mix protocol (P/N 4366128, Applied Biosystems). RT-PCR was carried out on the Applied Biosystems 7900HT Fast Real-Time PCR System using exon-boundary-specific TaqMan Gene Expression Assays (Applied Biosystems) (Table S1). Gene-expression data were analyzed using SDS 2.4.1 and Data Assist 3.0.1 software (Applied Biosystems).
Duquenne M., Folgueira C., Bourouh C., Millet M., Silva A., Clasadonte J., Imbernon M., Fernandois D., Martinez-Corral I., Kusumakshi S., Caron E., Rasika S., Deliglia E., Jouy N., Oishi A., Mazzone M., Trinquet E., Tavernier J., Kim Y.B., Ory S., Jockers R., Schwaninger M., Boehm U., Nogueiras R., Annicotte J.S., Gasman S., Dam J, & Prévot V. (2021). Leptin brain entry via a tanycytic LepR:EGFR shuttle controls lipid metabolism and pancreas function. Nature metabolism, 3(8), 1071-1090.
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