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| Objective: To explore the feasibility of quickly screening of gDNA library with PCR technique. Methods: We adopted porcine α-1,3GT cDNA fragment as the probe, used the primers synthesized by the specific sequence on cDNA to carry out α-1,3GT gene screening of porcine gDNA library by combining PCR and in situ plaque hybridization, and then performed enzymic digestion, southern blot, sequencing and fluorescence in situ hybridization for location. Results: After having finished one-time hybridization and one-time PCR, we obtained 7 positive monoclones with very strong signals, and each insert length of them is over 8kb, including the third intron. Moreover, 3 tested clones among them contain the third and fourth exons according to the sequencing results, and FISH mapped the inserts of the 3 clones to pig chromosome 1q2.10-q2.11. Conclusion: PCR could be applicable to the quick screening of DNA library and much simpler than the conventional in situ plaque hybridization only used. |
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| Keywords:genomic DNA library, PCR, hybridization |
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