以Chelex^R100作为介质、一步法从贮存骨髓涂片中提取DNA作PCR分析

Utilizing Chelex￣R 100 as A Medium for One-step Extraction of Dna From Stored Bone Marrow Smears for Use With PCR

利用螯合型离子交换树脂Ｃｈｅｌｅｘ~Ｒ１００作为介质、一步法从已贮存数年的骨髓细胞涂片中提取用作ＰＣＲ分析的ＤＮＡ。共提取已经或未经瑞氏染色的骨髓涂片２０张，平均每片ＤＮＡ收获率１．２±０．４μｇ。ＤＮＡ分子量为２～６ｋｂ大小，适合用于ＰＣＲ分析，但不能用于ＳｏｕｔｈｅｒｎＥｐ迹分析。本法具有简便、省时、明显减少因操作引起的标本间ＤＮＡ交叉污染的机会。此法有利于临床上收集大量的、不同病期的血液病标本，进行分子生物学研究和回顾性诊断。

Chelex￣R 100 is a chelating resin Byutilizing Chelex￣R 100 as a medium,DNA was extracted with a one-step procedure from 20 Wright-stained or unstained bone marrow smears which had been stored for several years.The average amount of DNA extracted from per slide was only 1.2±0.4μg with a size of 2-6kb.The DNA was partially degraded but suitable for PCR amplification,although unsuitable for Southern blot analysis.This method is simple.rapid and reduces dramatically the chance of operation-introduced DNA contamination or cross-contamination among samples.This technique can be widenes the scope of molecular analysis of hematological disorders to include the invstigation of anamnestic cases whose fresh tissue samples for DNA analysis are not available.