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CleanSEQ on the ABI 3100

pared to those observed using formamide and even higher if water elution is followed by the addition of mineral oil (Table 1). We propose that mineral oil prevents evaporation and blocks oxygen exchange with the sample, thereby slowing dye degradation and maintaining high signal to noise ratios. The increased signal to noise ratios produced using CleanSEQ with water elution allow extensive dilution of BigDye Terminators without affecting Phred20 read lengths (data not shown).


Extended Sample Stability
We assessed the stability of samples eluted in water and overlaid with mineral oil by comparing sequencing results from plates run immediately following CleanSEQ elution (fresh) versus samples reinjected 6 days later. Samples eluted in formamide were used as references, because they are known to be refractory to sample degradation. Both signal strength and Phred20 read length were unaffected by prior injection regardless of elution buffer (Figure 3). Signal to noise ratios were, however, far greater for samples eluted in water. These results clearly demonstrate that overlaying mineral oil stabilizes samples as well as formamide and allows flexible injection schedules.


Instrument Compatibility
Because much is proprietary about the manufacturing of ABI capillaries, Agencourt conducted thorough testing to confirm the compatibility of mineral oil and the 3100 capillary array. We injected samples eluted with water and overlaid with mineral oil in the same capillaries for 99 consecutive runs. Formamide samples were run in parallel in alternate capillaries as a base line of capil
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