實(shí)驗(yàn)?zāi)康?/strong>
對(duì)真菌細(xì)胞進(jìn)行破壁,提取其DNA。
實(shí)驗(yàn)原理
隨著分子生物學(xué)技術(shù)的進(jìn)展,PCR相關(guān)方法越來越多地被應(yīng)用到真菌學(xué)研究中來,而真菌DNA的提取是包括分子診斷在內(nèi)的所有實(shí)驗(yàn)的基礎(chǔ),簡(jiǎn)便快捷地提取真菌DNA,且獲得的DNA完整、純度高,對(duì)真菌分子生物學(xué)的研究有及其重要的意義。
真菌細(xì)胞壁以幾丁質(zhì)、葡萄糖為主構(gòu)成細(xì)胞壁骨架,而基質(zhì)則由多糖、甘露聚糖、糖蛋白、脂質(zhì)等填充。絲狀真菌堅(jiān)韌厚壁的構(gòu)成,需要較為繁瑣的破壁手段,故真菌DNA的提取較細(xì)菌或其他組織細(xì)胞難度更大。為保證真菌PCR技術(shù)及其相關(guān)分子生物學(xué)技術(shù)的順利開展,簡(jiǎn)便快捷地提取真菌DNA,且獲得的DNA完整、純度高,是必須解決的前提條件。
實(shí)驗(yàn)材料和器具
樣品:真菌類
儀器:高通量組織研磨儀(上海凈信,Tissuelyser-48)
耗材:離心管,研磨珠(上海凈信)
試劑:TE Buffer,DA Buffer,E Binding Buffer,Elution Buffer
實(shí)驗(yàn)步驟
需了解具體步驟,請(qǐng)致電廠家
實(shí)驗(yàn)結(jié)果
圖1 提取DNA的檢測(cè) M:100bp 標(biāo)準(zhǔn)參照物
圖2 提取DNA的PCR檢測(cè)結(jié)果 M:100bp 標(biāo)準(zhǔn)參照物
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