Table 1: Summary of results of cycle sequencing purification methods examined in this study.

TechniqueSequence length/qualityaCost, US $b
(per sample)
Time neededc
(per 96 samples)
AdvantagesDisadvantages

BioMag Carboxyl beads / 0.006–0.01525 minLow cost
Quick
Good results
Moderate potential for user error
Occasional blobs
Proprietary size exclusion columns / 2.2–2.63-4 hBest resultsCostly
Time consuming
Moderate potential for user error
Proprietary
in-well
technique
816 ± 1/99 ± 1*1.9437 minQuick
Good results
Low potential for user error
Costly
Short shelf life
Occasional blobs
Ethanol precipitation744 ± 35/87 ± 11**~0.0011 h 45 minLowest costLoss of amplicon
Low quality results
High potential for user error

Sequence length and quality scores for overlapping fragments were obtained directly from Sequencher 4.8 software (Gene Codes Corporation, Inc., Ann Arbor, MI). Prior to analyzing the data, raw sequences were trimmed by aligning all of the sequences of one reaction and determining the unusable part of the longest sequence and cutting all sequences at that base. Measurements and standard deviation are calculated using triplicate results of 4 different reactions (primers used were con2, con3, VP7R and 9con1L). Measurement based on *1 and **2 fewer sequences due to low quality of final sequence.
Cost per reaction was calculated by totaling the cost of consumables needed to clean one 10  L cycle sequencing reaction and does not account for the cost of equipment.
Time needed to purify 96 samples was measured from the moment the plate containing the cycle sequencing reactions was removed from the thermocycler after cycle sequencing to the time purification was completed and the 96-well septum was placed on the plate.