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[A rapid method for subtractive screening of hepatoma apoptotic cells cDNA library]
1Department of Pathology, Fourth Military Medical University, Xi'an, Shanxi. 710032 P. R. China mayg@plagh.com.cn
Objective:
To expedite the subtractive screening of human hepatoma apoptotic cells cDNA library.
Methods:
The method of subtractive hybridization combined with dot blot hybridization was adopted. First, minus cDNA probe was used to screen the cDNA library, the minus clones that did not hybridize with the minus cDNA probe were picked up as the source of the second grade dot blot hybridization. Secondly, two probes of the plus and minus cDNA were used to screen the clones;the clones that only hybridize with the plus cDNA probe were picked up as the source of the third grade dot blot hybridization.
Results:
Four clones were obtained and the lengths of the inserted cDNA fragments were about 1.5kb long.
Conclusion:
The results demonstrate this is a feasible, simple and quick method for subtractive screening of the cDNA library.