Total RNA from human HCC samples and cell lines was extracted using Beyozol (R0011, Beyotime, Shanghai) according to the standard guideline. The extracted total RNA was reverse-transcripted to the complementary DNA using BeyoRT cDNA synthesis kit (D7168L, Beyotime, Shanghai). The expression levels of SNHG6 and HIF1A were quantified using qPCR SYBR Green Master Mix (DRR041A, Takara, China). GAPDH was selected as the internal reference gene.
Beyort cdna synthesis kit
Manufactured by Beyotime
The BeyoRT cDNA synthesis kit is a reagent set designed for the reverse transcription of RNA into complementary DNA (cDNA). The kit includes the necessary components, such as reverse transcriptase enzyme, reaction buffer, and primers, to facilitate the conversion of RNA molecules into their DNA counterparts.
Automatically generated - may contain errors
4 protocols using beyort cdna synthesis kit
1
Quantifying SNHG6 and HIF1A in HCC
2
Quantitative Real-Time PCR Analysis of miRNA Expression
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Total RNA was extracted from the transfected cells using TRIzol reagent (Beyotime, Haimen, Jiangsu, China), according to the manufacturer’s protocols. The extracted RNA was reverse transcribed to cDNA using a BeyoRT™ cDNA synthesis kit (Beyotime), according to the manufacturer’s protocol. qRT-PCR was conducted using BeyoFast™ SYBR Green qPCR Mix (Beyotime) with an ABI 7500 Real-Time PCR System (Applied Biosystems; Thermo Fisher Scientific) according to the following protocol: initial step at 95°C for 60 seconds, followed by 95°C denaturation for 10 seconds, and 60°C annealing for 30 seconds (40 cycles). Expression levels of miR-384 were analyzed using the 2−ΔΔCt method with U6 snRNA as an internal control.14 (link) The primers used in this study are shown in Table 1 .
3
Quantification of SNHG6 and HIF1A
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Total RNA from human HCC samples and cell lines were extracted using Beyozol (R0011, Beyotime, Shanghai) according to the standard guidelines. The extracted total RNA was the reverse transcribed to complementary DNA using BeyoRT cDNA synthesis kit (D7168L, Beyotime, Shanghai). The expression levels of SNHG6 and HIF1A were quantitatively quantified using qPCR SYBR Green Master Mix (DRR041A, Takara, China). GAPDH was selected as the internal reference gene.
4
Ethical HCC Samples Collection and Analysis
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Ethics for HCC samples collecting and animal experiments HCC samples from patients were collected under the guidelines of the Ethics committee. And the animal experiments were conducted under the guidelines of the Animal care and use committee. All nude mice were purchased from Shanghai SLAC Laboratory Animal Co., Ltd, and housed under the room temperature and a 12-h light-dark cycle.
Total RNA extraction and quantitative PCR Total RNA from human HCC samples and cell lines was extracted using Beyozol (R0011, Beyotime, Shanghai) according to the standard guideline. The extracted total RNA was reverse-transcripted to the complementary DNA using BeyoRT cDNA synthesis kit (D7168L, Beyotime, Shanghai). The expression levels of SNHG6 and HIF1A were quanti ed using qPCR SYBR Green Master Mix (DRR041A, Takara, China). GAPDH was selected as the internal reference gene.
Total RNA extraction and quantitative PCR Total RNA from human HCC samples and cell lines was extracted using Beyozol (R0011, Beyotime, Shanghai) according to the standard guideline. The extracted total RNA was reverse-transcripted to the complementary DNA using BeyoRT cDNA synthesis kit (D7168L, Beyotime, Shanghai). The expression levels of SNHG6 and HIF1A were quanti ed using qPCR SYBR Green Master Mix (DRR041A, Takara, China). GAPDH was selected as the internal reference gene.
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