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保存好各个世代家禽血样和基因组DNA对于研究家禽各世代变化规律以及更好地监测家禽保种效果具有重要意义。试验分别抽取-20℃保存0.5年、1年、3年、5年、7年、8年和10年的狼山鸡全血基因组DNA样品,以及-70℃保存8年的狼山鸡DNA样品,采用核酸电泳和PCR检测,比较不同年份和不同温度保存DNA的效果。结果表明:DNA样品随着-20℃保存时间的延长,DNA越容易降解。-20℃保存10年的狼山鸡DNA降解率很高,条带模糊;-20℃保存8年和7年的DNA,降解也较严重;-20℃保存5年的狼山鸡DNA总体完好,条带相对清晰;-20℃保存3年、1年和0.5年的狼山鸡DNA条带清晰鲜明。-70℃保存8年的DNA条带清晰,无降解现象。降解的DNA浓度普遍显著低于不降解的DNA。DNA纯度均在1.8~1.9之间。降解和不降解的DNA用鸡内参引物PCR检测,均出现条带,说明DNA的部分降解不影响一般的PCR检测试验。本研究结果表明:-20℃保存5年以内DNA保存效果尚可,如果长期(5年以上)保存DNA,需-70℃超低温保存。
The preservation of blood samples and genomic DNA from poultry of all generations is of great importance for the study of the variation of each generation of poultry and the better monitoring of poultry preservation effect. The genomic DNA samples of Wolfhan chicken whole blood at -20 ℃ for 0.5 year, 1 year, 3 years, 5 years, 7 years, 8 years and 10 years were collected and the DNA samples of Wolfhan chicken preserved at -70 ℃ for 8 years , Using nucleic acid electrophoresis and PCR detection, comparing the different years and different temperature DNA preservation effect. The results showed that the DNA degraded more easily with the storage time of -20 ℃. DNA degradation rate of Wolfhan chicken stored at -20 ℃ for 10 years was very high, fuzzy bands were observed; DNAs stored at -20 ℃ for 8 and 7 years were more severely degraded; DNA of Wolfhan chicken kept at -20 ℃ for 5 years was generally intact , The band is relatively clear; -20 ℃ preserved 3 years, 1 year and 0.5 years Wolfshan chicken DNA bands clear and distinct. -70 ℃ DNA bands stored for 8 years clear, no degradation. Degraded DNA concentrations were generally significantly lower than non-degraded DNA. DNA purity are between 1.8 ~ 1.9. Degraded and non-degraded DNA was detected by PCR with chicken reference primers, showing bands, indicating that partial degradation of DNA did not affect normal PCR assays. The results of this study show that DNA preservation can be preserved within 5 years of storage at -20 ℃. If DNA is stored for a long time (more than 5 years), it needs to be stored at -70 ℃ for cryopreservation.