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[DNA sequencing of human ciliary neurotropic factor gene by silver staining method]
1Institute of Medical Sciences, Nanjing Railway Medical College, Nanjing, 210009 P. R. China.
Objective:
To introduce a non-radioactive protocol for DNA sequence analysis which employs a silver staining procedure
Methods:
DNA template is amplified by Taq DNA polymerase to generate sequence ladder, No labeled nucleotide or primer is involved in the thermal-cycling sequencing reaction. A sensitive silver staining procedure is employed to visualize bands in sequencing gel.
Results:
This silver staining method is used for DNA sequence analysis of human cilliary neurotrophic factor (hCNTF) gene. The resulted band resolution is comparable to radioactive sequencing method, and data can be obtain within less than 2 hours after sequencing reaction.
Conclusion:
Silver staining is a rapid, inexpensive and reliable method for DNA sequence analysis.