QuantiTect Primer Assays (Qiagen) and Brilliant II SYBR Green QRT-PCR 1-step system (Agilent Technologies) were used following the manufacturer’s instructions. GAPDH was used as loading control. The following primers were used (Qiagen): GAPDH (QT00079247), MMP-1 (QT00014581), MMP-2 (QT00088396), MMP-9 (QT00040040), MMP-13 (QT00001764), MMP-14 (QT00001533) and JAK1 (QT00050225). However, primer sequences are not provided by Qiagen, as stated in their website: ‘Sequences of the QuantiTect Primer Assays are not provided. Approximate location of primers within a specific gene can be viewed on the Product Detail pages retrieved via our GeneGlobe data base.’
Brilliant 2 sybr green qrt pcr 1 step system
Manufactured by Agilent Technologies
The Brilliant II SYBR Green QRT-PCR 1-step system is a laboratory equipment product from Agilent Technologies. The core function of this product is to facilitate one-step quantitative reverse transcription polymerase chain reaction (QRT-PCR) analysis using SYBR Green as the fluorescent reporter dye.
Automatically generated - may contain errors
Lab products found in correlation
Quantitect primer assay, by Qiagen (4 mentions)
Gapdh, by Qiagen (3 mentions)
Mmp 1, by Qiagen (2 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Abi prism 7900ht sequence detection system, by Thermo Fisher Scientific (1 mentions)
Quantitect primer, by Qiagen (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
4 protocols using brilliant 2 sybr green qrt pcr 1 step system
1
Quantitative RT-PCR of Matrix Metalloproteinases
2
Isolation and qRT-PCR Analysis of RNA
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RNA was isolated using the RNeasy Mini Kit (Qiagen), according to manufaturer’s instructions, and QuantiTect Primer Assays, purchased from Qiagen, were used. The Brilliant II SYBR Green QRT-PCR 1-Step system was used according to the manufacturer’s instructions (Agilent Technologies). The reactions were analysed on an ABI PRISM 7900HT Sequence Detection System (Applied Biosystems).
3
Quantitative RT-PCR of Matrix Metalloproteinases
4
Quantifying LIMK, MKL, and Myosin Expression
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RNA was isolated using TriZol (Life technologies). For experiments comparing expression in parental vs BRAFi-resistant cells (A375- and Colo829-derivatives), resistant cells were cultured with 1 μM PLX4720 and sensitive cells with equivalent volume of DMSO for 24 hr. QuantiTect Primer Assays (Qiagen) and Brilliant II SYBR Green QRT-PCR 1-step system (Agilent Technologies) with 100 ng RNA were used following the manufacturer’s instructions. GAPDH was used as loading control. The following QuantiTect Primers were used (Qiagen): GAPDH (QT00079247), LIMK1 (QT00008680), LIMK2 (QT00084357), MKL1 (QT00067921), MKL2 (QT00010115), MYH9 (QT00073101), MYL9 (QT00072268), MYL12A (QT01665741), MYL12B (QT00075264), ROCK1 (QT00034972), ROCK2 (QT00011165). Primer sequences are not provided by Qiagen, as stated in their website: ‘Sequences of the QuantiTect Primer Assays are not provided. Approximate location of primers within a specific gene can be viewed on the Product Detail pages retrieved via our GeneGlobe data base.’
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