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The myth of automated, high‐throughput two‐dimensional gel analysis
Author(s) -
Clark Brittan N.,
Gutstein Howard B.
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700709
Subject(s) - replicate , computer science , throughput , software , matching (statistics) , spots , process (computing) , pattern recognition (psychology) , artificial intelligence , statistics , chemistry , mathematics , programming language , operating system , wireless
Many software packages have been developed to process and analyze 2‐D gel images. Some programs have been touted as automated, high‐throughput solutions. We tested five commercially available programs using 18 replicate gels of a rat brain protein extract. We determined computer processing time, approximate spot editing time, time required to correct spot mismatches, as well as total processing time. We also determined the number of spots automatically detected, number of spots kept after manual editing, and the percentage of automatically generated correct matches. We also determined the effect of increasing the number of replicate gels on spot matching efficiency for two of the programs. We found that for all programs tested, less than 3% of the total processing time was automated. The remainder of the time was spent in manual, subjective editing of detected spots and computer generated matches. Total processing time for 18 gels varied from 22 to 84 h. The percentage of correct matches generated automatically varied from 1 to 62%. Increasing the number of gels in an experiment dramatically reduced the percentage of automatically generated correct matches. Our results demonstrate that these 2‐D gel analysis programs are not automatic or rapid, and also suggest that matching accuracy decreases as experiment size increases.