Cell identification

細(xì)胞鑒定

近年來，大量研究表明 STR 基因分型方法是進(jìn)行細(xì)胞交叉污染和性質(zhì)鑒定的最有效和準(zhǔn)確的方法之一，STR基因分型應(yīng)用于細(xì)胞鑒定已被ATCC等機(jī)構(gòu)強(qiáng)烈推薦。美國的ATCC 細(xì)胞庫、德國的DSMZ細(xì)胞庫以及日本的 JCRB細(xì)胞庫等為STR 分型提供了各細(xì)胞株的數(shù)據(jù)供比對。

STR基因位點由長度為2～7個堿基對的短串連重復(fù)序列組成，這些重復(fù)序列廣泛存在于人類基因組中，可作為高度多態(tài)性標(biāo)記，被稱為細(xì)胞的DNA指紋，其可通過PCR（聚合酶鏈?zhǔn)椒磻?yīng)）來檢測。STR基因座位上的等位基因可通過擴(kuò)增區(qū)域內(nèi)重復(fù)序列的拷貝數(shù)的不同來區(qū)分，在毛細(xì)管電泳分離之后可通過熒光檢測來識別。隨后通過一定的計算方法，即可根據(jù)所得的STR分型結(jié)果與專業(yè)的細(xì)胞STR數(shù)據(jù)庫比對從而推算出樣品所屬的細(xì)胞系或可能的交叉污染的細(xì)胞系名稱。

實驗方案：

收樣
細(xì)胞DNA
提取
DNA擴(kuò)增
STR位點和
性別基因檢測
STR分型圖譜
和數(shù)據(jù)庫比對

樣本要求：

活細(xì)胞/細(xì)胞團(tuán)/凍存株數(shù)量要求：≥10^6個；
DNA送樣標(biāo)準(zhǔn)是：DNA濃度>50ng/μl，體積≥20μl；
活細(xì)胞一般為20℃以上常溫寄送，細(xì)胞團(tuán)藍(lán)冰寄送，DNA一般也是藍(lán)冰寄送，除非客戶特殊要求干冰出庫，凍存株使用干冰寄送。

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參考文獻(xiàn)：

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