Tissue (100 mg) or cells were ground to powder in liquid nitrogen, 1 ml of TRIzol was added to fully lyse the cells, and RNA was extracted. The PrimerScript Reverse Transcription Kit (RR047A, Takara, Shiga, Japan) was used for cDNA preparation and the TB Green™ Premix Ex Taq (RR420A, Takara) was used for qPCR. Relative gene expression levels were analyzed using the 2−ΔΔCt method. interleukin (IL)-6, or interferon (IFN)-γ in culture supernatants of LUAD cell line were quantified using IL-6/IFN-γ Quantikine ELISA kit (R&D Systems, CA, United States) according to the manufacturer’s instructions.
Primerscript reverse transcription kit
Manufactured by Takara Bio
Sourced in Japan, United States
The PrimerScript Reverse Transcription Kit is a laboratory tool designed for the reverse transcription of RNA into complementary DNA (cDNA). It contains the necessary reagents and enzymes to efficiently perform this fundamental step in various molecular biology applications.
Automatically generated - may contain errors
2 protocols using primerscript reverse transcription kit
1
RNA Extraction and Gene Expression Analysis
2
Quantitative RT-PCR Analysis of LUAD
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Total RNA was extracted from LUAD tissue samples and cultured cells using Trizol reagent (Invitrogen, USA) according to the manufacture’s protocol. The RNA was reverse transcribed into cDNA using PrimerScript Reverse Transcription Kit (Takara, RR047A) and amplified by qRT-PCR with TB Green™ Premix Ex Taq (Takara RR420A) in an ABI 7500 System (Applied Biosystems, Foster City, CA, USA). Actin housekeeping gene acted as the internal control. The 2−ΔΔCt method quantified the expression of the target gene. The primers used in this study were acquired from Invitrogen Shanghai Sangon Biotech (Invitrogen, China). The primers sequences were as follows: LINC00551 Forward: 5ʹ-TCAGCAGCCTTCAGTTGGAG-3ʹ Reverse: 5ʹ-AGGCTCTGTTCAGCTGGTTC-3ʹ; c-Myc Forward: 5ʹ-GGCTCCTGGCAAAAGGTCA-3ʹ Reverse: 5ʹ-CTGCGTAGTTGTGCTGATGT-3ʹ; Actin Forward: 5ʹ-AGCGAGCATCCCCCAAAGTT-3ʹ Reverse: 5ʹ- GGGCACGAAGGCTCATCATT-3ʹ; PKM2 Forward: 5ʹ-ATGTCGAAGCCCCATAGTGAA-3ʹ Reverse: 5ʹ-TGGGTGGTGAATCAATGTCC-3ʹ.
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