摘要
目的探讨恒温ＤＮＡ自动测序中几种因素对测序结果的影响。方法对ＤＮＡ纯度、浓度和测序过程中所用水的质量对ＤＮＡ自动测序结果的影响进行了回顾性研究。结果当ＤＮＡ纯度较低时，核苷酸曲线的信噪比降低，或多个核苷酸峰重叠而判读困难；使用低浓度ＤＮＡ时，所能判读的核苷酸长度明显短于较高浓度时；测序反应过程中和配制测序胶时所用的纯水水质较差时，核苷酸峰可变宽、变平，或出现双峰，影响判读结果。结论ＤＮＡ纯度和浓度以及测序反应中及配制测序胶时所用水的质量对测序结果有明显影响。提示恒温ＤＮＡ自动测序时须使用高纯度的ＤＮＡ和高质量的纯水；若要获得较长ＤＮＡ片段的序列。
Objective To investigate the influence of several factors on isothermal automated DNA sequencing. Methods The effects of DNA purity and DNA concentration,as well as the quality of water used during sequencing on automated DNA sequencing results were retrospectively analyzed. Results When the DNA was not pure enough,the nucleotide curve showed lower ratio of signal to noise,or compression would occur,leading to difficulty in correctly judging the nucleotides;the readable length of nucleotides was shorter when low concentration of DNA was used;when the quality of water used in sequencing reaction and polyacrylamide gel solution was not high enough,the peaks of nucleotides became wider and flatter,or double peaks would occur,resulting in unreadable sequences. Conclusion DNA purity and concentration as well as quality of water used in sequencing reaction and polyacrylamide gel solution had apparent influence on isothermal automated DNA sequencing.This suggests that high purity of DNA and high quality of water should be used in isothermal automated DNA sequencing;if longer DNA sequence needs to be obtained,DNA concentration should not be too low.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
1998年第4期238-241,共4页
Chinese Journal of Medical Genetics
关键词
DNA
自动测序
测序
DNA纯度
水质
Isothermal automated DNA sequencing DNA purity Water quality
