( nerated using the RoboCycler ...)Automated Fluorescent-Tag Cycle Sequencing

nerated using the RoboCycler Gradient 96 temperature cycler with Hot Top Assembly, showing read length to 745 bases. figure 2 panel B shows the results generated from the cycle sequencing reactions run on the GeneAmp PCR System 9600 cycler, showing read length to 720 bases. The sequence read from reactions cycled on the RoboCycler Gradient 96 temperature cycler required 2 hours and was 1% inaccurate or unassignable. The sequence read from reactions cycled on the GeneAmp PCR System 9600 cycler (using the manufacturers protocol) took over 20 minutes longer than the reactions cycled with the RoboCycler unit. In addition, the sequence generated from reactions cycled on the GeneAmp system was 2% inaccurate or unassignable. The intensities of the fluorescent-tag signals were more uniform for the reactions performed on the RoboCycler Gradient 96 temperature cycler with Hot Top Assembly than for the reactions performed on the GeneAmp PCR System 9600 cycler. This difference in uniformity may have caused the difference in accuracy of base recognition between reactions performed on the two instruments.

Conclusions

With its unmatched well-to-well temperature accuracy, the RoboCycler Gradient 96 temperature cycler with Hot Top Assembly performs fluorescent-dye cycle sequencing reactions accurately and reproducibly. The new Hot Top Assembly for both gradient and nongradient RoboCycler 96 temperature cyclers eliminates the need for oil or wax overlays on reactions during PCR cycle sequencing, which makes loading samples onto automated DNA sequencing instruments quick and easy.

REFERENCES

1. Adams, M.D., Fields, C., and Venter, J.C., eds. (1994) In Automated DNA Sequencing and Analysis. Academic Press, San Diego, Calif
   '"/>

Source:


Page: All 1 2 3 4 5
Related biology technology :

1. Purified Single-Stranded M13 DNA for Automated Sequencing
2. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
3. The DIG System Nonradioactive Automated High-Throughput In Situ Hybridization: a Powerful Tool for Functional Genomics Research
4. Agilent 2100 Bioanalyzer Automated Analysis System
5. Automated RNA Amplification for Arrays
6. Quantitation of DNA for Automated Sequencing Using the VersaFluor Fluorometer
7. Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A
8. Total RNA Purification from Cultured Cells Using the ABI PRISM 6700 Automated Nucleic Acid Workstation and Total RNA Lysis Reagents
9. DNA Damage and Repair - Quantification of Sub-Cellular Events Using Automated Confocal Imaging
10. Small-Molecule Quantitation Using Automated Single-Shot Averaging MALDI-TOF Mass Spectrometry
11. Automated Dissolution of Prednisone with the MultiDose G3