Sequence verification as quality-control step for production of cDNA microarrays.
To generate cDNA arrays in our core laboratory, we amplified about 2300 PCR products from a human, sequence-verified cDNA clone library. As a quality-control step, we sequenced the PCR products immediately before printing. The sequence information was used to search the GenBank database to confirm the identities. Although these clones were previously sequence verified by the company, we found that only 79% of the clones matched the original database after handling. Our experience strongly indicates the necessity to sequence verify the clones at the final stage before printing on microarray slides and to modify the gene list accordingly.
Taylor
E
E
University of Texas, M.D. Anderson Cancer Center, Houston, TX, USA.
Cogdell
D
D
Coombes
K
K
Hu
L
L
Ramdas
L
L
Tabor
A
A
Hamilton
S
S
Zhang
W
W
D015723:Gene Library; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards*; D016133:Polymerase Chain Reaction; D011786:Quality Control; D017422:Sequence Analysis, DNA / Q000592:standards