A user-friendly alternative to formaldehyde-based DNA silver-staining method on polyacrylamide gels.
We have developed a practical, cost-effective and user-friendly protocol to meet the needs of nucleic acids research, particularly in respect of DNA detection on polyacrylamide gels. In this method, the most commonly used alkaline formaldehyde developer in DNA silver stain, which does harm to operator, is first replaced by glucose in alkaline borate buffer. In addition, the effects of six reducing sugars on the quality of DNA visualization were investigated. Consequently, the optimal protocol using glucose takes about 45 min to complete all the procedures, with a detection limit of 5 pg of single DNA band on polyacrylamide gels, was developed. The results indicate that this user-friendly and economic protocol could be a good choice for routine use in DNA visualization on polyacrylamide gels.
He
Hong-Zhang
HZ
Zhejiang Provincial Key Laboratory of Biopharmaceuticals, Wenzhou Medical College, Wenzhou, PR China.
Cong
Wei-Tao
WT
Jiang
Cheng-Xi
CX
Pu
Jie
J
You
Wei-Jing
WJ
Gao
Hong-Chang
HC
Zhu
Zhong-Xin
ZX
Jin
Li-Tai
LT
Li
Xiao-Kun
XK
D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005557:Formaldehyde; D005947:Glucose / Q000737:chemistry; D057230:Limit of Detection; D016622:Silver Staining / Q000379:methods*
D005557:Formaldehyde; D004247:DNA; D005947:Glucose