Simple Two-Color Base-Calling Schemes for DNA Sequencing Based on Standard Four-Label Sanger Chemistry

$29.00 plus tax (Refund Policy)

Buy Article:

Abstract:

Two base-calling schemes for DNA sequencing are evaluated. Both are based on data collected from two broad-band emission channels derived from either one- or two-excitation channels. Standard four-dye Sanger reaction products are used in conjunction with capillary electrophoretic separation in a polymer matrix. Data acquisition is compatible with high light-throughput imaging and minimal data storage. In one scheme, commercial chromatographic software provides peak recognition and peak heights. The peak-height ratios from the two channels provide base-calling accuracies of 99.3% and 97.1% through 330 bp and 350 bp, respectively. In another scheme, ratiograms are derived from the two channels. The resulting step-like functions permit the calling of bases even when successive peaks are not resolved. The base-calling accuracy is 99% through 340 bp. Because of simplicity in implementation, either scheme should be readily applicable to high-speed, high-throughput DNA sequencing in capillary arrays.

Keywords: Base calling; Capillary electrophoresis; DNA sequencing; Spectroscopic imaging

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/0003702953965470

Affiliations: Department of Chemistry and Ames Laboratory-USDOE, Iowa State University, Ames, Iowa 50011

Publication date: October 1, 1995

More about this publication?
Related content

Tools

Favourites

Share Content

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more