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Construction and application of human liver tissue cDNA library

Jun Sheng, Hongwei Zhu, Dapeng Zhao, Zhi‐Wu Wang, Mei Chen, Zhu Liu, Hua Zhou, Xuanjun Wang, Wei An

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Abstract

After the total RNA of liver tissue was extracted by CsCl_(2)-GTC ultracentrifugation, mRNA was isolated by corresponding kit and used for synthesizing the first chain of cDNA in the present of reverse transcriptase and the second chain in the present of E.coli DNA polymerase I and RNase H, using M13M4 Olygo d(T)_(8) (containing the EcoR I site) as primer. In order to verify the reliability of synthesized cDNA library, the authors designed several pairs of primers used for the amplification of TPO and IGF-1 genes by PCR. The result showed that TPO and IGF-1 genes were successfully amplified and segregated. The results of restriction enzyme digestion and DNA sequencing indicated that the sequences of the 2 genes completely agreed with those reported in published documents.

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What this paper is about

After the total RNA of liver tissue was extracted by CsCl_(2)-GTC ultracentrifugation, mRNA was isolated by corresponding kit and used for synthesizing the first chain of cDNA in the present of reverse transcriptase and the second chain in the present of E.coli DNA polymerase I and RNase H, using M13M4 Olygo d(T)_(8) (containing the EcoR I site) as primer. In order to verify the reliability of synthesized cDNA library, the authors designed several pairs of primers used for the amplification of TPO and IGF-1 genes by PCR. The result showed that TPO and IGF-1 genes were successfully amplified and segregated. The results of restriction enzyme digestion and DNA sequencing indicated that the sequences of the 2 genes completely agreed with those reported in published documents.

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Available abstract

After the total RNA of liver tissue was extracted by CsCl_(2)-GTC ultracentrifugation, mRNA was isolated by corresponding kit and used for synthesizing the first chain of cDNA in the present of reverse transcriptase and the second chain in the present of E.coli DNA polymerase I and RNase H, using M13M4 Olygo d(T)_(8) (containing the EcoR I site) as primer. In order to verify the reliability of synthesized cDNA library, the authors designed several pairs of primers used for the amplification of TPO and IGF-1 genes by PCR. The result showed that TPO and IGF-1 genes were successfully amplified and segregated. The results of restriction enzyme digestion and DNA sequencing indicated that the sequences of the 2 genes completely agreed with those reported in published documents.

Key concepts: Complementary DNA, Reverse transcriptase, Primer (cosmetics), Molecular biology, cDNA library, RNase H, Gene, Biology

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Construction and application of human liver tissue cDNA library — Research Paper | ScholarLens