Assessing the reliability of amplified RNA used in microarrays: a DUMB table approach.
A certain minimal amount of RNA from biological samples is necessary to perform a microarray experiment with suitable replication. In some cases, the amount of RNA available is insufficient, necessitating RNA amplification prior to target synthesis. However, there is some uncertainty about the reliability of targets that have been generated from amplified RNA, because of nonlinearity and preferential amplification. This current work develops a straightforward strategy to assess the reliability of microarray data obtained from amplified RNA. The tabular method we developed, which utilises a Down-Up-Missing-Below (DUMB) classification scheme, shows that microarrays generated with amplified RNA targets are reliable within constraints. There was an increase in false negatives because of the need for increased filtering. Furthermore, this analysis method is generic and can be broadly applied to evaluate all microarray data. A copy of the Microsoft Excel spreadsheet is available upon request from Edward Bearden.
Bearden
Edward D
ED
Department of Geriatrics, University of Arkansas for Medical Sciences, and Department of Biostatistics, Arkansas Children's Hospital, Little Rock, Arkansas 72205, USA. beardenedwardd@uams.edu
Simpson
Pippa M
PM
Peterson
Charlotte A
CA
Beggs
Marjorie L
ML
D052160:Benzothiazoles; D045744:Cell Line, Tumor; D019295:Computational Biology / Q000379:methods*; D003959:Diamines; D005188:False Negative Reactions; D006801:Humans; D015233:Models, Statistical; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry; D009930:Organic Chemicals / Q000494:pharmacology; D011804:Quinolines; D012313:RNA / Q000737:chemistry*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D052160:Benzothiazoles; D003959:Diamines; D009841:Oligonucleotides; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D012313:RNA