DNA fragments containing full-length TUG1 or the negative control sequences were amplified by PCR, using primers containing t7. The resulting fragments were cloned into a plasmid vector GV394 (Genechem, China), which had been linearized by XhoI digestion. Reverse transcription of biotin-labeled RNA was performed using T7 RNA polymerase (Takara Biomedical Technology, Japan) and biotin RNA labeling mixture (Roche, USA). RNA was extracted using the RNeasy Mini Kit (Qiagen, USA), treated with RNase-free DNase I (Roche, USA), then subjected to qRT-PCR as previously described above.
Gv394
Manufactured by Genechem
Sourced in China
The GV394 is a laboratory equipment designed for general use. It serves as a versatile tool for various applications within a research or clinical setting. The core function of the GV394 is to provide a controlled environment for tasks requiring temperature regulation and precise monitoring.
Automatically generated - may contain errors
6 protocols using gv394
1
Cloning and Quantifying TUG1 Transcripts
2
Biotinylated CASC2 Probe Synthesis
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The biotinylated DNA probe complementary to CASC2 was amplified by PCR using T7-containing primer (Roche, Basel, Switzerland) and then inserted into GV394 (Genechem, Shanghai, China). The resultant plasmids DNAs were linearized using restriction enzyme XhoI. Biotin-labeled RNAs were transcribed with Biotin RNA Labeling Mix (Roche) and T7 RNA polymerase (Takara). The products were treated with RNase-free DNase I (Roche) and purified with the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). RNA were extracted for RT-qPCR analysis.
3
Biotin-labeled LINC00337 RNA Production
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The DNA fragment with the full-length LINC00337 or NC sequence was amplified using a primer with T7 and cloned into GV394 from Genechem (Shanghai, China). The restriction enzyme XhoI was used for the linearization of DNA. Next, T7 RNA polymerase (Takara) and Biotin RNA Labeling Mix (Roche, China) were used for reverse transcription of biotin-labeled RNAs that underwent reverse transcription. Thereafter, the products received DNase I (RNase-free, Roche) treatment and purification using the RNeasy Mini Kit (Qiagen, USA), and the extracted RNAs were used for qRT-PCR assessment.
4
CASC2 Sequence Cloning and RNA Synthesis
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The DNA fragment with the full length CASC2 sequence or negative control sequence was PCR amplified using a T7-containing primer and then cloned into GV394 (Genechem, Shanghai, China). The resultant plasmids DNAs were linearized using restriction enzyme XhoI. Biotin-labeled RNAs were reversely transcribed using Biotin RNA Labeling Mix (Roche, USA) and T7 RNA polymerase (Takara Biomedical Technology). The products were treated with RNase-free DNase I (Roche, USA) and purified with the RNeasy Mini Kit (Qiagen, USA) and RNA were extracted for real-time PCR.
5
LINC00337 Transcription Protocol
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The DNA fragment with the full length LINC00337 or NC sequence received PCR ampli cation using a primer with T7 and was cloned into GV394 from Genechem, Shanghai, China). Restriction enzyme XhoI was utilized for linearization of DNAs. Next, T7 RNA polymerase (Takara) and Biotin RNA Labeling Mix from Roche (China) were employed for reverse transcription of biotin-labeled RNAs underwent reverse transcription. Thereafter, the products received DNase I (RNase-free, Roche) treatment and puri cation using the RNeasy Mini Kit (Qiagen, USA), and the extracted RNAs were employed for qRT-PCR assessment.
6
Biotinylated MALAT1 RNA Probe Protocol
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The biotinylated DNA probe complementary to MALAT1 RNA was amplified by PCR using a T7-containing primer and subcloned into the plasmid vector GV394 (GeneChem). The resultant plasmid was linearized by restriction enzyme XhoI. Biotin-labeled RNAs were reversely transcribed using biotin RNA Labeling Mix and T7 RNA polymerase (all from Roche, Indianapolis, Ind). Finally, the bound RNAs were purified with the RNeasy Mini Kit (Qiagen, Hilden, Germany) and then extracted for further evaluation by qRT-PCR analysis.
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