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Quantitect assays

Manufactured by Qiagen
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Sourced in United States

Quantitect assays are a range of ready-to-use kits designed for sensitive and reliable gene expression analysis. The core function of these assays is to provide optimized reagents and protocols for quantitative real-time PCR (qPCR) to measure the expression levels of target genes.

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Lab products found in correlation

Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Biomark 96.96 dynamic array platform, by Standard BioTools (1 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) Reliaprep rna cell miniprep system, by Promega (1 mentions) Sybr green, by Thermo Fisher Scientific (1 mentions) Abi7900ht pcr machine, by Thermo Fisher Scientific (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Direct zol rna kit, by Zymo Research (1 mentions) Moloney murine leukaemia virus reverse transcriptase, by Promega (1 mentions) Sybr green jumpstart taq readymix, by Merck Group (1 mentions) Quantitect, by Qiagen (1 mentions) Chromo4 real time pcr detector, by Bio-Rad (1 mentions) Quantitect sybr green pcr master mix, by Qiagen (1 mentions)

Close spelling variants of this product

Specific QuantiTect Primer Assays SYBR Green-based QuantiTect Primer assays Quantitect gene expression assays Quantitect SYBR-green RT-PCR assay kit Mm-Il18-1-SG QuantiTect primer assay QuantiTect primer assay for mouse Tlr9 Dr_bdnf_1_SG QuantiTect Primer Assay Hs_GAPDH_2_SG QuantiTect Primer Assay Hs_RRN18S_1_SG QuantiTect Primer assay 18S QuantiTect Primer Assay

5 protocols using quantitect assays

1

Quantitative RNA expression analysis

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1 microgram total RNA was treated with 1U DNaseI (Promega 9PMIM610) and cDNA prepared with SuperscriptIII reverse transcriptase (Invitrogen) and random primers. Targets were quantified with 1x Fast Sybr (ABI) and 1x Quantitect assays (Qiagen) or Taqman assays by the delta CT method using B2m as normalizer (Supplementary Table S16).
Uribe-Lewis S., Carroll T., Menon S., Nicholson A., Manasterski P.J., Winton D.J., Buczacki S.J, & Murrell A. (2020). 5-hydroxymethylcytosine and gene activity in mouse intestinal differentiation. Scientific Reports, 10, 546.
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2

Quantitative RT-PCR Analysis of Gene Expression

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For Figure 2, 1 μg total RNA was treated with 1U DNaseI (Promega 9PMIM610) and cDNA prepared with SuperscriptIII reverse transcriptase (Invitrogen) and random primers. Targets were quantified with 1× Fast Sybr (ABI) and 1× Quantitect assays (Qiagen) by the standard curve method using serial dilutions of cDNA template from Jeg3 cells and normalised to B2M. For supplementary Figure 3, 1 μg RNA was reverse transcribed using Quantitect reverse transcription kit (Qiagen) following manufacturer’s instructions. Real-time PCR used the 96.96 Biomark Dynamic Array platform (Fluidigm) and Taqman assays (ABI) following manufacturer’s instructions (Fluidigm). Fold change between normal and tumour was calculated by the delta Ct method using B2M as the normaliser. All expression assays are listed in Additional file 1.
Uribe-Lewis S., Stark R., Carroll T., Dunning M.J., Bachman M., Ito Y., Stojic L., Halim S., Vowler S.L., Lynch A.G., Delatte B., de Bony E.J., Colin L., Defrance M., Krueger F., Silva A.L., ten Hoopen R., Ibrahim A.E., Fuks F, & Murrell A. (2015). 5-hydroxymethylcytosine marks promoters in colon that resist DNA hypermethylation in cancer. Genome Biology, 16(1), 69.
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3

Quantifying BCR-ABL Fusion Gene Expression

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Total RNA was isolated using the Direct-zol™ RNA Kit (Zymo Research) or ReliaPrep™ RNA Cell Miniprep System (Promega), and synthesis of cDNA was performed (Thermo Fisher). Quantitative real time PCR analyses were performed on ABI7900HT PCR machine (Applied Biosystems) using Quantitect assays (Qiagen) and SYBR Green (Applied Biosystems). The expression of ABL1 and the fusion BCR-ABL (m-bcr; e1-a2) were measured using TaqMan Gene expression assays (Hs01104728_m1 ABL1 and Hs03024844_ft BCR-ABL, respectively) from Applied Biosystems. Relative quantification was calculated using 2−ΔΔCT equation.
Abdelrasoul H., Vadakumchery A., Werner M., Lenk L., Khadour A., Young M., El Ayoubi O., Vogiatzi F., Krämer M., Schmid V., Chen Z., Yousafzai Y., Cario G., Schrappe M., Müschen M., Halsey C., Mulaw M.A., Schewe D.M., Hobeika E., Alsadeq A, & Jumaa H. (2020). Synergism between IL7R and CXCR4 drives BCR-ABL induced transformation in Philadelphia chromosome-positive acute lymphoblastic leukemia. Nature Communications, 11, 3194.
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4

Quantitative RT-PCR Analysis of Gene Expression

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Measurement of mRNA expression was carried out essentially as described
previously(31). Briefly, complementary DNA was prepared from 1 µg of the same RNA as
used for the microarray using Moloney-murine leukaemia virus reverse transcriptase
(Promega). Quantitative RT-PCR was performed in a total reaction volume of 10 µl with SYBR
Green JumpStart Taq ReadyMix (Sigma) and QuantiTect primer assays (Table 1; Qiagen). mRNA levels were determined by the standard curve
method(32) and normalised to glyceraldehyde 3-phosphate dehydrogenase
(GAPDH) expression, which was found to be unaffected in all three cell
lines. All samples were analysed in duplicate.

Quantitative RT-PCR primer assays*

TranscriptQuantiTect assay
ASMTLQT00097720
FOLR1QT01000853
GAPDHQT00079247
MXRA8QT00202790
NMBQT00066745
PHF5AQT00080360
SLC19A1QT00014742
SLC25A4QT00048279
SLC46A1QT00067515
UBE2J1QT00029855

* Primers were QuantiTect assays purchased from Qiagen. Primer sequences were not
available from the company.

Price R.J., Lillycrop K.A, & Burdge G.C. (2016). Folic acid induces cell type-specific changes in the transcriptome of breast cancer cell lines: a proof-of-concept study. Journal of Nutritional Science, 5, e17.
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5

Quantitative Analysis of Immune Markers

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The levels of transcripts were measured by real-time PCR using human genes QuantiTect Assays and QuantiTect Hs_GAPDH Assay (Qiagen, Valencia, CA, USA) as a normalizer. The following genes were assessed: FOXP3, CD152 (CTLA-4), CD28, CD80 (B7-1), CD86 (B7-2), CD40, CD154 (CD40L), and CRP (C-reactive protein). Real-time PCR was performed in duplicate in 20 μL, using the QuantiTect SYBR Green PCR Master Mix (Qiagen) applying the manufacturer's instructions, and conducted in the Chromo4 Real-Time PCR Detector (BIO-RAD, USA). A standard curve construction was generated by a series of four dilutions of cDNA of the control group sample in reaction to the house-keeping gene, GAPDH. On the basis of these curves, the levels of total chosen gene transcripts were calculated after their normalization to GAPDH. The value of CT was determined by the first cycle number at which fluorescence was higher than the set threshold value.
Pawlowski P., Wawrusiewicz-Kurylonek N., Eckstein A., Reszec J., Luczynski W., Johnson K., Kretowski A., Bakunowicz-Lazarczyk A., Gorska M., Szamatowicz J., Chyczewski L, & Mysliwiec J. (2015). Disturbances of Modulating Molecules (FOXP3, CTLA-4/CD28/B7, and CD40/CD40L) mRNA Expressions in the Orbital Tissue from Patients with Severe Graves' Ophthalmopathy. Mediators of Inflammation, 2015, 340934.
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