Extraction Of Genomic DNA From Mice Tissue And Cloning Of 3' Flanking Of MWAP Gene

This research studied the function of NaCl and SDS and the relationship of proteinase K and EDTA in lysing mice tissue and the effects of some other factors on mice tissue lysis through lysing mice tissue and extracting genomic DNA. And discussed the mechanism of mice tissue lysis based on these studies. The 3′flanking of mice whey acid protein gene was cloned which is about 1750 bp and sequenced and analysed in this research. The related results of this research are as following:1.NaCl have a big effect on mice tissue lysis through lysing the same kind of mice tissue in the same kind of lysis buffer additional NaCl of different concentration. Mice tissue can′t be lysed well in a lysis solution of NaCl of low concentration. The lysis solution of 0.1mol.L-1 NaCl can lyse mice tissue well and is adopted in many lysis solutions.2.EDTA have an effect on the activity of proteinase K by comparison of the concentration of EDTA and PrK in lysis solutions and their lysis effect. Much PrK is desired in lysis solutions of EDTA of high concentration.3.SDS can inactive nuclease and protect DNA well by comparison of the concentration of EDTA in different lysis solutions and the quality of DNA extracted by the same method. And the concentration of EDTA in lysis solutions could not be too high.4.Lysis tempreture has an effect on tissue lysis in this research. None other than the tempreture which make SDS and PrK work well at the same time can make tissue lysed well. Too high tempreture is worse than useless. The lysis tempreture in common use is 50-55℃.5.Pre-treatments such as splicing tissue and squeezing tissue have not good effect on tissue lysis, and sometimes they may delay tissue lysis while the pretreatment is incorrect.6.Thinking over the above results it can be found that lysis components may diffuse in tissue through intercelluler gelatinization stroma and lyse cells and NaCl may feature in the formation and maintenance of gelatified state of intercelluler stroma.7.Some methods of DNA extraction and purification were compared in this research and find that the method of glass rod twining and the method of centrifugation column are practical in mice genomic DNA extraction and purification.8.The 3′flanking of mice whey acid protein gene was cloned successfully which is about...