LncRNA biomarkers for diagnosing human lung adenocarcinoma and human colorectal cancer
A biomarker, colorectal cancer technology, applied in the field of LncRNA biomarkers for the diagnosis of human lung adenocarcinoma and human colorectal cancer
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Embodiment 1
[0040] 1 Clinical sample collection:
[0041] A total of 113 pairs of frozen fresh tissue specimens of early lung adenocarcinoma and their paired normal lung tissues were collected from Zhongshan Hospital Affiliated to Fudan University during 2011-2014. All patients gave informed consent to participate in the scientific research.
[0042] 2 LncRNA quantitative PCR experiment:
[0043] 2.1 RNA extraction and quantitative PCR
[0044] Total RNA was extracted from frozen tissues using Trizol, and RNA was quantified using a microvolume spectrophotometer.
[0045] 2.2 Internal reference genes and primers
[0046] Internal reference gene selection GAPDH, GAPDH and a set of primer sequences of LncRNA biomarkers provided by the present invention are shown in Table 1:
[0047] Table 1 Primer sequences used in quantitative PCR
[0048]
[0049]
[0050] 2.3 Reverse transcription reaction
[0051] Use the SuperScriptⅢFirst Strand Synthesis System kit for reverse transcription...
Embodiment 2
[0076] 1 Clinical sample collection:
[0077] 18 pairs of fresh frozen tissue specimens of colorectal cancer and their paired normal colorectal tissues were collected from Ruijin Hospital Affiliated to Shanghai Jiaotong University in 2013. All patients gave informed consent to participate in the scientific research.
[0078] 2 LncRNA quantitative PCR experiment:
[0079] 2.1 RNA extraction and quantitative PCR
[0080] Total RNA was extracted from frozen tissues using Trizol, and RNA was quantified using a microvolume spectrophotometer.
[0081] 2.2 Internal reference genes and primers
[0082] GAPDH was selected as the internal reference gene, and the primer sequences of the internal reference gene and each LncRNA biomarker are shown in Table 1 in Example 1.
[0083] 2.3 Reverse transcription reaction, real-time quantitative PCR reaction
[0084] The specific methods, steps and reaction system of the reverse transcription reaction and the real-time quantitative PCR reactio...
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