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KMID : 0545120010110050890
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Journal of Microbiology and Biotechnology
2001 Volume.11 No. 5 p.890 ~ p.894
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Rapid Isolation of Genomic DNA from Normal and Apoptotic Cells Using Magnetic Silica Resins
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Park, Jee Sun
Park, Jung Hyun/Na, Shin Young/Choe, Soo Young/Choi, Sang Nam/You, Kwan Hee
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Abstract
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The isolation of genomic DNA from mammalian cells is usually performed by cell lysis followed by protein digestion, extraction, and finally, ethanol precipitation of the chromosomal DNA. However, in the case of large sample numbers or when fast processing is required or when only small amounts of starting materials are available, such conventional methods are not efficient and are cumbersome to be applied. Some alternative methods have been described as well as having commercial DNA isolation kits to be available, nevertheless, there is room left for much improvement. In the present study, a novel method is introduced, where it simplifies conventional protocols by omitting some time-consuming steps such as protease incubation or DNA precipitation and its resuspension. Using paramagnetic silica resins, the genomic DNA was purified over a magnetic field, and the bound DNA was eluted with a low-salt buffer. The fidelity and effectiveness of this novel method was determined by using normal and apoptotic cells as a starting material and then compared to other protocols. The high speed and convenience along with its high efficiency in detecting apoptotic chromosomal DNA will prove this method to be an improved alternative in the isolation of genomic DNA from mammalian cells.
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