Comparison and modification of rRNA sequencing methods

Wang, R.F.; Cao, W.W.; Slavik, M.F.

Biotechniques 11(4): 438; 440

1991


ISSN/ISBN: 0736-6205
PMID: 1793573
Document Number: 377466
In this study, modification of two methods of RNA sequencing resulted in more definitive sequencing bands. In one method of sequencing, the bands of lane A and lane G sometimes were not clear. Modifications of this method by changing the concentrations of ddATP and ddGTP resulted in the bands of lane A and lane G becoming more readable. Although a second sequencing method was found to have clearer bands than the first method, and the bases immediately downstream from the primer binding site could be read by using r-32P-labeled primer, the bands on the top of lane A still were not clear. Modifications of this second method by changing the ddATP/dATP ratio resulted in the bands of lane A becoming much clearer.

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