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- W2337609872 abstract "【Objective】 The study was to construct a long-length cDNA library of skin tissue for Xinji Fine-wool Sheep to seek for genes related to wool trait.【Method】 The library was constructed by SMART technique using CreaterTM SMARTTM cDNA library construction kit.The total RNA was extracted from sheep skin tissue using Biozol isolation reagent.Single-strand cDNA was synthesized using SuperscriptTM Ⅱ reverse transcriptase,and double-strand cDNA was synthesized and amplified by long-distance PCR.The PCR products were purified by QIAquickTM PCR Purification kit.After digestion with SfiⅠand size fractionation using CHROMA SPIN-400 columns,ds cDNA was ligated to the SfiⅠ-digested,dephosphorylated pDNR-LIB vector.The ligation mixture was transformed into E.coli DH5α by thermal shock.【Result】 The cDNA library contained 4.2×105 independent clones with DNA inserts of 0.5-3.0 kb.The recombination rate was 100% and the average length of inserts was 1.3 kb.【conclusion】 The capacity of this library can meet the requirement for gene screening and provide a foundation to isolate specific genes in skin for Xinji Fine-wool Sheep." @default.
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- W2337609872 date "2010-01-01" @default.
- W2337609872 modified "2023-10-06" @default.
- W2337609872 title "Construction of a long-lenth cDNA library for Xinji Fine-wool Sheep skin tissue by use of SMART technique." @default.
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