Total RNA from tissues and cells was extracted with TRIzol reagent (Invitrogen, Carlsbad, CA, USA), and cDNAs were synthesized according to the manufacturer’s instructions by using TaqMan MultiScribe Reverse Transcriptase (Applied Biosystems, Foster City, CA, USA). qRT-PCR analysis was performed using an ABI Prism 7900-HT Sequence Detection System (96-well, Applied Biosystems). Primers were designed and synthesized by RiboBio (Guangzhou, China). The primers used in qRT-PCR are listed in Supplementary Tables S1 & S2 . The relative expressions of lncRNAs and mRNAs were normalized to GAPDH, and miRNAs were normalized to U6. The experiments were performed in triplicate.
Abi prism 7900 ht sequence detection system 96 well
Manufactured by Thermo Fisher Scientific
The ABI Prism 7900-HT Sequence Detection System (96-well) is a real-time PCR instrument designed for high-throughput gene expression analysis. It features a 96-well format and supports multiple quantitative PCR chemistries.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (4 mentions)
Taqman multiscribe reverse transcriptase, by Thermo Fisher Scientific (2 mentions)
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
Cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Rnase free dnase set, by Qiagen (1 mentions)
4 protocols using abi prism 7900 ht sequence detection system 96 well
1
Quantitative Analysis of RNA Expression
2
c-FLIPL Expression Analysis by qRT-PCR
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Total cellular RNA was isolated with TRIzol reagent (Invitrogen). Samples were treated with DNase using the RNase-free DNase Set (Qiagen) during the total RNA isolation. First-strand complementary DNA (cDNA) was synthesized using the cDNA Synthesis kit (Thermo Fisher Scientific) according to the manufacturer's instructions. ABI prism 7900-HT sequence detection system (96-well, Applied Biosystems) was used to perform quantitative real-time PCR (RT-PCR) analysis. For RT-PCR, the following primers were used:
c-FLIPL: 5'-AGAACCTGGCTGCACCTAAC-3'(forward),
5'-GAGAAGGTCAAACCGCCTCA-3'(reverse).
GAPDH: 5'-CGAGATCCCTCCAAAATCAAGTGGGG-3'(forward),
5'-ACACGTTGGCAGTGGGGACAC-3'(reverse).
3
Quantitative RNA Expression Analysis
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Total RNA from tissues and cells was extracted with TRIzol reagent (Invitrogen, Carlsbad, CA, USA), and cDNA was synthesized according to the manufacturer’s instructions by using a RevertAid™ First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, Waltham, MA, USA). RT-qPCR analysis was performed using an ABI Prism 7900-HT Sequence Detection System (96-well, Applied Biosystems). Primers were designed and synthesized by RiboBio (Guangzhou, China). The primers used in RT-qPCR are listed in Supplementary Tables 1 and 2. The relative expression of lncRNAs was normalized to that of GAPDH, and miRNAs were normalized to U6.
4
Quantifying PD-L1 mRNA Expression in Cells
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To quantify PD-L1 mRNA expression, total RNA was extracted from cells with TRIzol reagent (Invitrogen), and cDNA was synthesized using TaqMan MultiScribe Reverse Transcriptase (Applied Biosystems, Foster City, CA) according to the manufacturer's instructions. Quantitative realtime PCR (RT-PCR) analysis was performed using an ABI Prism 7900-HT Sequence Detection System (96-well, Applied Biosystems). For RT-PCR, the following primers were used for the amplification of PD-L1, forward primer 5′-CCTACTGGCATTTGCTGAACGCAT-3′ and reverse primer 5′-ACCATAGCTGATCATGCAGCGGTA-3′. β-Actin: 5′-TCC TGTGGCATCCACGAAACT-3′ (forward) and 5′-GAAGCA TTTGCGGTGGACGAT-3′ (reverse). The experiments were triplicated. The relative expression of PD-L1 was normalized to β-actin expression. Different PD-L1 mRNA levels of other cell lines were relative to that of Beas-2B cells.
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