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Magnetic Microspheres: Synthesis and Application in Genomic DNA Extraction

Author: ZhangLiMing
Tutor: WanQianHong
School: Tianjin University
Course: Pharmaceutical Analysis
Keywords: Magnetic separation Magnetic microspheres Polyethylene glycol Calf thymus DNA The Ephron Wunderlich adsorption model Saccharomyces cerevisiae Corn kernels
CLC: R944
Type: Master's thesis
Year: 2005
Downloads: 318
Quote: 5
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Abstract


With the rapid development of the life sciences and bio-engineering , magnetic separation technology with its simple and fast separation applications , suitable for a variety of sample matrix , easy automation and miniaturization characteristics is becoming an indispensable means of the field of life sciences . The laboratory using a template method and chemical coprecipitation method three different matrix types of magnetic microspheres and silica gel as adsorbent , adsorption solution of polyethylene glycol (PEG) and sodium chloride (NaCl) , research calf thymus deoxyribonucleic acid (DNA) in the adsorption behavior of the silanol - based solid-phase carrier . The experimental results show that the adsorption of the solution was composed of 20 % ( w / v ) PEG and 2.0 mol / L NaCl , and elution time of 10 min under the conditions of up to 80% , the recovery of DNA . Similar to the the magnetic microspheres extraction efficiency with silicone , but by eliminating the need for centrifugation steps to shorten the time of preparation of nucleic acid template , conducive to automated operation . In order to study the adsorption mechanism of the DNA on the magnetic microspheres , we further studied the adsorption solution consisting of PEG and NaCl , the DNA is adsorbed to a solid phase carrier on the adsorption isotherm , the study found that the adsorption of DNA can be used Ephron Delhi Greek adsorption model to describe the correlation coefficient can be more than 0.994 . Polishing method pulverized mammalian tissue calf thymus and plant tissue , corn kernels , Cell Lysates broken cells to silanol surface of magnetic microspheres as a solid phase carrier for the magnetic separation of genomic DNA extraction to obtain a fragment of about 20 kb, A 260 / A 280 greater than 1.77 high purity DNA . The magnetic microspheres are used in a wild Saccharomyces cerevisiae genomic DNA extraction . Using two different methods for partial and complete lysis of the yeast cell wall , to obtain a fragment of about 5 kb and 20 kb respectively , and A 260 / A 280 in 1.77-1.89 between the DNA fragment. For the preparation of magnetic microspheres after surface modification with a silylating agent , the various surface functional groups of magnetic microspheres used to calf thymus extract of corn kernels , and wild yeast genomic DNA . Of magnetic microspheres with the surface of the silanol , to obtain better results.

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