MMP16 mRNA levels were analyzed using a real-time PCR assay. Total RNA was isolated from tissue samples or cultured cells using TRIzol (Invitrogen), and reversely transcripted to cDNA with PrimeScript™ RT Master Mix (Perfect Real Time) kit (RR036A, Takara) based on the manufacturer's instruction. Real-time PCR was performed with the SYBR Green master mix kit using an ABI-7900 thermal cycling instrument (Applied Biosystems). GAPDH gene was amplified as an endogenous control. Primers were as follows: GAPDH-F, 5′- GCA CCGTCAAGGCTGAGAAC-3′, GAPDH-R, 5′-TGGTG AAGACGCCAGTGGA-3′; MMP16-F, 5′- GGACAGAAA TGGCAGCACAAGC -3′, MMP16-R, 5′- CATCAAAGG CACGGCGAATAGC -3′.
Abi 7900 thermal cycling instrument
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI-7900 is a thermal cycling instrument designed for quantitative real-time PCR (qRT-PCR) applications. The core function of the ABI-7900 is to precisely control and regulate the temperature of samples during the thermal cycling process required for qRT-PCR experiments.
Automatically generated - may contain errors
2 protocols using abi 7900 thermal cycling instrument
1
Quantifying MMP16 mRNA Expression
2
LINC00511, miR-625-5p, and GSPT1 Expression Analysis
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LINC00511, miR-625-5p, and GSPT1 mRNA in the patients’ tissues and cells were measured by qRT-PCR. Total RNA in each tissue and cell sample was collected using TRIzol (Thermo Fisher Scientific). To synthesize complementary DNA (cDNA), the same amount of each RNA sample was subjected to reverse transcription reaction. The reverse transcription reaction was conducted using a PrimeScript RT reagent kit (Takara Bio, Kusatsu, Japan), and all operations were carried out according to the manufacturer’s instructions. The expression level of the above 3 genes was then evaluated by qRT-PCR using a SYBR Premix Ex Taq kit (Takara Bio). An ABI-7900 thermal cycling instrument was provided by Applied Biosystems (Waltham, MA, USA). With glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control, the 2−ΔΔCT method was used to analyze the relative expression of genes.
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