Total RNA was isolated from tumor tissues, adjacent tissues, and serum samples. Reverse transcription was conducted with a TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA) to synthesize the first chain of cDNA. Then, RT-PCR reaction was performed with β-actin as internal control in the Applied Biosystems 7900 Fast Real-Time PCR system (Applied Biosystems, Foster City, CA, USA). The expression of lncRNA-D16366 was evaluated using the comparative cycle threshold (CT).
Applied biosystems 7900 fast real time pcr system
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The Applied Biosystems 7900 Fast Real-Time PCR system is a high-performance real-time PCR instrument designed for rapid and precise gene expression analysis. It features a 96-well block format and supports a wide range of fluorescent chemistries and applications.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (10 mentions)
Taqman universal master mix 2, by Thermo Fisher Scientific (6 mentions)
Nuclease free water, by Thermo Fisher Scientific (6 mentions)
High capacity cdna reverse transcription kit with rnase inhibitor, by Thermo Fisher Scientific (5 mentions)
Taqman gene expression assay 20, by Thermo Fisher Scientific (4 mentions)
Trizol, by Thermo Fisher Scientific (4 mentions)
Sybr premix ex taq, by Takara Bio (4 mentions)
Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (3 mentions)
Rneasy mini kit, by Qiagen (3 mentions)
Biometra tadvanced thermocycler, by Analytik Jena (2 mentions)
Taqman gene expression assay 20x, by Thermo Fisher Scientific (2 mentions)
Sybr premix ex taq kit, by Takara Bio (2 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions)
Primescript rt reagent kit, by Takara Bio (2 mentions)
Biometra tadvanced, by Analytik Jena (1 mentions)
Primescript rt reagent kit, by Qiagen (1 mentions)
Sybr green 1 assay, by Takara Bio (1 mentions)
Trizol agent, by Thermo Fisher Scientific (1 mentions)
Primescrip rt reagent kit, by Takara Bio (1 mentions)
Sybr green premix, by Takara Bio (1 mentions)
28 protocols using applied biosystems 7900 fast real time pcr system
1
Quantification of lncRNA-D16366 Expression
2
Gene Expression Analysis of Brain Tumor Cell Lines
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RNA expression assays were performed for SLC12A5 (Hs00221168_m1), SLC12A2(Hs00169032_m1), CDH1 (Hs01023894_m1), CDH2 (Hs00983056_m1) and GAPDH (Hs02786624_g1) genes. According to the manufacturer’s instructions, reverse transcription was performed with the High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor (Applied Biosystems, Waltham, MA, USA) in a 20 µL reaction volume containing 50 ng RNA using the Biometra TAdvanced thermal cycler (Analytik Jena AG, Jena, Germany). The synthesized copy DNA (cDNA) was stored at 4 °C until use or at −80 °C for a longer time. Real-time polymerase chain reaction (PCR) was performed using an Applied Biosystems 7900 Fast Real-Time PCR System (Applied Biosystems, Waltham, MA, USA). The reactions were run in triplicate with 4 µL of cDNA template in a 20 µL reaction volume (10 µL of TaqMan Universal Master Mix II, no UNG (Applied Biosystems, Waltham, MA, USA), 1 µL of TaqMan Gene Expression Assay 20× (Applied Biosystems, Waltham, MA, USA), 5 µL of nuclease-free water (Invitrogen, Carlsbad, CA, USA)), with the program running at 95 °C for 10 min, followed by 45 cycles of 95 °C for 15 s, 50 °C for 2 min and 60 °C for 1 min.
The control and 24 h TMZ-treated groups (n = 6 per group) were tested for SLC12A5, SLC12A2, CDH1 and CDH2 expression.
The control and 24 h TMZ-treated groups (n = 6 per group) were tested for SLC12A5, SLC12A2, CDH1 and CDH2 expression.
3
Quantification of Osteoclast Gene Expression
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Total RNA was isolated from cultured OCPs or osteoclasts using an RNeasy Mini Kit (Qiagen, Hilden, Germany) and reverse‐transcribed to cDNA using a PrimeScript RT Reagent Kit (Qiagen), according to the manufacturer's instructions. Gene expression was measured using an Applied Biosystems 7900 Fast Real‐Time PCR System (Applied Biosystems, Foster City, CA) with the TaqMan Gene Expression Assays software program. Also used were PCR primers specific to TRACP (Acp5, Mm00475698_m1), cathepsin K (Ctsk, Mm00484039_m1), nuclear factor of activated T cells, cytoplasmic, calcineurin dependent 1 (Nfatc1, Mm00479445_m1), dendritic cell‐expressed seven transmembrane protein (Dcstamp, Mm01219007_m1), PR domain‐containing 1 (Blimp1, Prdm1, Mm00476128_m1), B cell leukemia/lymphoma 6 (Bcl6, Mm00477633_m1), RANK (Tnfrsf11a, Mm00437132_m1), and Cx3cr1 (Mm00436454_m1). mRNA levels were normalized to that of the housekeeping gene hypoxanthine phosphoribosyltransferase (Hprt, Mm01545399_m1). PCR products were quantified using the comparative cycle threshold (Ct) method (2 − ΔΔCt).(22 )
4
RNA Extraction and qRT-PCR Analysis
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Total RNA was extracted from the collected specimens using Trizol agent (Invitrogen, Carlsbad, CA) following the manufacturer's instructions. DNaes I was used to treat the residual DNA in RNA samples. The concentration and quality of the RNA samples were detected by UV absorbance (A260/A280) and 1% agarose gel electrophoresis, respectively. The first chain of cDNA was compounded through a Prime Scrip RT reagent kit (Takara Biotechnology Co, Ltd). Real-time polymerase chain reaction (RT-PCR) was performed with SYBR Green I assay (Takara) in the Applied Biosystems 7900 Fast Real-Time PCR system (Applied Biosystems, Foster City, CA). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) acted as internal control. The relative expression of RASSF10 was calculated with 2−ΔΔCt method. The sequences of primers used in this study were as followed. RASSF10: forward 5′-CCATGACCCAGGAGAAACAG-3′; reverse 5′-GCTGGCGAATTGTGTGGTC-3′. GAPDH: forward 5′-CATGAGAAGTATGACAACAGCCT-3′; reverse 5′-AGTCCTTCCACGATACCAAAGT-3′.
5
RT-qPCR Assay for Slc12a2 and Glpdh
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RNA expression assay was performed for Slc12a2 (Rn00582505_m1) and Glpdh (Rn01775763_g1) genes. High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor (Applied Biosystems, USA) was used for reverse transcription reaction in 20 μl reaction volume containing 50 ng of total RNA incubated at 25°C for 10 min, transcripted at 37°C for 120 min, and terminated by heating at 85°C for 5 min using Biometra TAdvanced thermocycler (Analytik Jena AG, Germany). The synthesized cDNA was stored at 4°C until use or at -20°C for longer time. The Real-time PCR was run in triplicate with 4 μl of cDNA template in a 20 μl reaction volume (10 μl of TaqMan Universal Master Mix II, no UNG (Applied Biosystems, USA), 1 μl of TaqMan Gene Expression Assay 20x (Applied Biosystems, USA), and 5 μl of Nuclease-Free Water (Invitrogen, USA) with the program running at 95°C for 10 min, followed by 40 cycles of 95°C for 15 s and 60°C for 1 min. the reaction was performed using an Applied Biosystems 7900 Fast Real-Time PCR System (Applied Biosystems, USA).
6
Skin Cancer miR-148a Expression
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Total RNA was extracted from the patients with skin cancer and healthy controls with TRIzol (Invitrogen, Carlsbad, CA, USA). The reverse transcription was made to synthesize the first chain of cDNA with TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA). Then RT-PCR reaction was performed in the Applied Biosystems 7900 Fast Real-Time PCR system (Applied Biosystems, Foster City, California, USA). GAPDH was taken as the internal control. The relative expression quantification of miR-148a at mRNA level was evaluated by comparative cycle threshold (CT) method.
7
RNA Expression Profiling of SLC Transporters
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RNA expression assays were performed for SLC12A2 (Hs00169032_m1), SLC12A5 (Hs00221168_m1), SLC5A8 (Hs00377618_m1) and GAPDH (Hs02786624_g1) genes. According to the manufacturer’s instructions, reverse transcription was performed with the High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor (Applied Biosystems, Waltham, MA, USA) in a 20 µL reaction volume containing 50 ng RNA using the Biometra TAdvanced thermal cycler (Analytik Jena AG, Jena, Germany). The synthesized copy DNA (cDNA) was stored at 4 °C until use, or at −80 °C for a longer time. Real-time polymerase chain reaction (PCR) was performed using an Applied Biosystems 7900 Fast Real-Time PCR System (Applied Bio-systems, Waltham, MA, USA). The reactions were run in triplicate with 4 µL of cDNA template in a 20 µL reaction volume, 10 µL of TaqMan Universal Master Mix II, no UNG (Applied Biosystems, Waltham, MA, USA), 1 µL of TaqMan Gene Expression Assay 20× (Applied Biosystems, Waltham, MA, USA), 5 µL of nuclease-free water (Invitrogen, Carlsbad, CA, USA), with the program running at 95 °C for 10 min, followed by 45 cycles of 95 °C for 15 s, 50 °C for 2 min and 60 °C for 1 min.
The control and 24 h VPA-treated groups (n = 6 per group) were tested for SLC12A2, SLC12A5, SLC5A8 and GAPDH expression.
The control and 24 h VPA-treated groups (n = 6 per group) were tested for SLC12A2, SLC12A5, SLC5A8 and GAPDH expression.
8
Quantitative Analysis of miR-494 and lncRNA-XIST Expression
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Total RNA from 10 mm spinal cord segment containing the injury epicenter was isolated using TRIzol (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instructions. After reverse transcription, cDNA was amplified by using SYBR-Green Premix (Takara, Otsu, Japan). The expression of miR-494 and lncRNA-XIST in tissue was, respectively, normalized to the expression of U6 and GAPDH. RT-qPCR was performed using the Applied Biosystems 7900 Fast Real-Time PCR system (Applied Biosystems, Foster City, CA, USA). The data were analyzed by ΔCt method. The sequences of primers were purchased from Guangzhou RiboBio Co. Ltd.: lncRNA-XIST forward 5′-CGGGTCTCTTCAAGGACATTTAGCC-3′, and reverse 5′-GCACCAATACAGAGGAATGGAGGG-3′; GAPDH forward, 5′-GAAGATGGTGATGGGATTTC-3′, and reverse, 5′-GAAGGTGAAGGTCGGAGT-3′; miR-494 forward, 5′-TGACCTGAAACATACACGGGA-3′ and reverse, 5′-TATCGTTGTACTCCACTCCTTGAC-3′; U6 forward, 5′-AAAGACCTGTACGCCAACAC-3′ and reverse, 5′-GTCATACTCCTGCTTGCTGAT-3′.
9
Profiling lncRNA-AK001085 in Ankylosing Spondylitis
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TRIzol®reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) was used to isolated the total RNA from the serum of patients with AS and healthy controls. Reverse transcriphase synthesized the first chain of cDNA with SuperScript™ III (Invitrogen Life Technologies). Then RT-PCR reaction was performed in the Applied Biosystems 7900 Fast Real-Time PCR system (Applied Biosystems, Foster City, California, USA) with a internal control of β-actin. Comparative cycle threshold (CT) method was utilized for calculating the relative expression of lncRNA-AK001085.
10
Quantifying Lnc-LSAMP-1 and LSAMP in NSCLC
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Total RNA from 170 paired NSCLC tissues and 11 cell lines were extracted using TRIzol reagent (Invitrogen, Carlsbad, California, USA). The total RNA was then transcribed to cDNA using commercial kits according to the manufacturer’s instructions (TaKaRa, Japan). RT-qPCR reaction (DBI, Germany) was performed in the Applied Biosystems 7900 Fast Real-Time PCR system (Applied Biosystems, CA, USA). β-actin was used as the endogenous control. The primers were synthesized by Sangon Biotech Ltd (Shanghai, China). The primer sequences used for RT-qPCR were presented in Additional file 4 : Table S1. The 2−ΔΔCT was used to demonstrate the expression levels of Lnc-LSAMP-1 and LSAMP. All the experiments were conducted in triplicate.
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