Base on the instruction of the tissue RNA extraction kit (Tiangen Biotechnology Co., Ltd., Beijing, China), total RNA was extracted. Then complementary DNA (cDNA) was synthesized using PrimeScript RT Reagent kit (TaKaRa, Dalian, China) with 1 μg RNA, and the qRT-PCR reaction was carried out by SYBR Green qPCR Master Mix (TaKaRa). The sequence of primers were as follows: TP73-AS1 forward, 5ʹ-GCTCCGTGAACCAACTCG-3ʹ, and reverse, 5ʹ-GCTGCCAAGGGAACTCT-3ʹ; miR-494-3p forward, 5ʹ- GGGTGAAACACACACGGGAA-3ʹ, and reverse, 5ʹ- GGCAGGTCCGAGGT-3ʹ; EIF-5A2 forward, 5ʹ-TTCCAGCACTTACCCTT-3ʹ, and reverse, 5ʹ-TTCCCCTCTATTTTTG-3ʹ; GAPDH forward, 5ʹ-TCGACAGTCAGCCGCATCTTCTTT-3ʹ, and reverse, 5ʹ-ACCAAATCCGTTGACTCCGACCTT-3ʹ; U6 forward, 5ʹ-CTCGCTTCGGCAGCACA-3ʹ, and reverse, 5′-AACGCTTCACGAATTTGCGT-3ʹ. GAPDH and U6 were used as inside parameters to normalize the data, and the mRNA expression level was calculated using 2−ΔΔ Ct.
Tissue rna extraction kit
Manufactured by Tiangen Biotech
Sourced in China
The Tissue RNA Extraction Kit is a laboratory tool designed to efficiently extract and purify total RNA from various tissue samples. It utilizes a guanidinium thiocyanate-based lysis buffer and silica-based membrane technology to capture and wash the RNA, resulting in high-quality RNA suitable for downstream applications such as RT-qPCR, RNA sequencing, and gene expression analysis.
Automatically generated - may contain errors
2 protocols using tissue rna extraction kit
1
Quantitative Analysis of RNA Expression
2
Gastric Cancer Cell Line Silencing
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Gastric cancer cell line MGC-803 was purchased from American Type Culture Collection (ATCC). RhoA-silenced A6-B9 cells and COX-2-silenced D7-B8 were prepared by ourselves. Tissue RNA extraction kit was purchased from Tiangen Biotech (Beijing) Co., Ltd. Reverse transcription kit was purchased from Fermentas. Semi-quantitative PCR kit was purchased from ThermoFisher. Rabbit polyclonal antibodies against RhoA and COX-2 were purchased from Santa Cruz. Goat anti-rabbit polyclonal antibodies were purchased from Shanghai Pufei Biotechnology Co., Ltd. Rabbit immunohistochemistry staining kit was purchased from Shanghai Yanji Biotechnology Co., Ltd.
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