[The influence of several factors on automated DNA sequencing].
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.


CONCLUSIONS
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.
Xiao
C
C
Department of Medical Genetics, The First Affiliated Hospital, West China University of Medical Sciences, Chengdu, Sichuan, 610041 P. R. China. zszg@wcums.edu.cn
Zhang
S
S
Wu
H
H
Xia
Q
Q
D004247:DNA / Q000032:analysis; D017422:Sequence Analysis, DNA / Q000379:methods*; D014867:Water
D014867:Water; D004247:DNA