3种禽血抗凝剂效果及其对DNA提取质量的影响

通过3种不同血液抗凝剂对禽血抗凝,提取新鲜的和经冷冻保存一段时间后的抗凝血DNA,运用紫外分光法、凝胶电泳和PCR扩增技术检测提取的DNA样品质量,探讨不同抗凝剂的抗凝效果及其对提取DNA质量的影响。结果表明:3种抗凝血提取的DNA纯度和浓度相近。新鲜抗凝血比冷冻保存一段时间后的抗凝血提取的DNA质量好。ACD抗凝血在冷冻一段时间后出现凝血块,EDTA抗凝血液放置时间过长也影响抗凝效果,但两者在加入裂解液后则可长时间保存。75%乙醇抗凝剂抗凝效果较好,可长时间保存,适合远距离采血。由3种抗凝禽血提取的DNA分子片段完整,能满足常规分子生物学试验研究。

Effect of Three Anticoagulants on Chicken's Blood and the Quality of DNA Extraction

Three chicken blood anticoagulants were used and the quality of the DNAs extracted from fresh and frozen/melt anticoagulant blood samples were detected with ultraviolet absorption,Ago-Gel electrophoresis and PCR amplification.The results demonstrated that concentration and purity of the DNAs extracted were comparable among the 3 anticoagulant treatments.The quality of the DNA extracted from fresh anticoagulant blood sample was better than that from frozen/melt ones.75% alcohol was suitable for long distance sample collection,and was the best one in storing the anticoagulant bloods for long time in homogenization liquid status after frozen/melt compared with the ACD and EDTA treatments.All the DNAs extracted from the 3 kinds of anticoagulant bloods can be applied for further normal manipulation of molecular biologic analysis.