Simple method for DNA extraction from pancreatic juice for PCR amplification assays

被引:14
作者
Müller, P
Jesnowski, R
Liebe, S
Rolfs, A
Löhr, M
机构
[1] Univ Rostock, Div Gastroenterol, Dept Med, D-18057 Rostock, Germany
[2] Univ Rostock, Dept Neurol, D-18057 Rostock, Germany
关键词
PCR; pancreatic juice; DNA; ras; DNA preparation;
D O I
10.1385/IJGC:25:1:39
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Conclusion: Preparation of DNA from pancreatic juice for subsequent polymerase chain reaction (PCR) is difficult, but manageable, The protocol presented offers a simple and fast solution. This method might be applicable to other complicated samples, such as saliva, wound secretions, or stool washings. Background: Of all the biological samples used for PCR amplification, pancreatic juice is the most problematic because of the presence of potential inhibitory substances and the amount of nucleases. This demands a DNA preparation procedure that is suitable for routine diagnostic PCR, and is therefore efficient and safe. This is particularly true for pancreatic juice obtained during routine endoscopy. Methods: We describe here a simple method utilizing modified phenol/chloroform extraction and precipitation directly from native pancreatic juice suitable for diagnostic PCR applications, such as oncogenes. Results: DNA could be prepared in quantitative amounts from routine endoscopic specimens. DNA could also be prepared from samples kept several days at room temperature.
引用
收藏
页码:39 / 43
页数:5
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