Evidence of cross-hybridization artifact in expressed sequence tags (ESTs) on cdna microarrays
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1 E -b (EST) DA D H, Sb D P Rb O D M F L W P S, R S C G, b, Ab W -b - DA DA - (T). T (T), (T), (EST) DA RA. A b DA EST - T b -. O (T) b -b RA. T b DA b b b EST (T). I, DA (T) DA b. K: DA M, E-T S, EST, C-b, P(T), P(A),. ITRODUCTIO S DA,,, /. T ( ), ( ). M, b. A, b () b C, H, Sb, P, O D, F, W, S, S, G,. C M U, D S U Pb, S Pb H, H G U Pb, S M, D C M U, D P I H M C, U W F
2 () b. T, b, (.., ). O, : b b,,, b b b. I b. S, DA (EST) (T). W -. RESULTS D. W DA, - DA U Pb (P. b F. b ) - (C/C) b S M Db b (B. ). W. A, b b. P (T) b b. W b RA (T). I (P.) % b b T. A (EST). I, % b (T). U b, (T). E. I b,. A (b), ( ). O, (T). O b b, (T). T b, -b, RA. E (T). W (T). T (), (), (). I,, : (T). I b (T), T b,. I, W - DA. T CBI b AI T b C.
3 -, (T) (-) (-); b. O, -, b. E b (T). T b (T), b (T). F C =,,..., b : V = C V (X ) C V (X ) C (T) V (X ) (.., ). W V b b C,, T-, (,, C V, ). T,, b C b (T). F - DA, b (T). H, -, b b (T). E b (T). W b b (T) b. E b, C ( b T, ). F, (,,, b ). F, b (T). T, - DA, b ( P./F. ), (T) ( ). T b, T- b b b. T. T b. I, b b b b b ( ) (T), b. T : H : F, (T). H : I. P-. Tb -. A α =.,, H, DA DA. - α =. -. T DA (T). T b E. H, -.
4 - DA b (T) b (T) -. TZD b b Tb : E - - DA, - DA, -(C/C) DISCUSSIO E. E (EST) (- b) DA b. T, RA,, (.., ), / (.., ). T RA -b (T). T DA,, -, b b [B]. B b, EST (%)[Wb & L], b. EST. W DA, RA -A. T - (T) b GB. T EST - (T) b -b. S DA b b b b- () DA, b b DA A T b EST DA (T). T b. O b, b, - b -- b. H, b. W b b -b. S DA b-, -b b b -A (T). I,, b b -A DA. T b EST b,. W (T), CBI GB,. T-. A - DA b -. I -, -b b ( ) b,. O -.
5 H, - EST (T), b -b. L -b b. T b. T, (.., C C) b, -- b (T). I. E EST b. A -b b. W b DA. I, - EST (T). S. B, -b DA, b AOVA,,,. F,, b b. I F & D (), b b b. METHODS. TZD- T DA RA TL - (TZD)(P,. b ). C T (I C., Cb, CA) RA. RA /. DA b RA. T RA./L (T) C. R X - b (I C., Cb, CA).L (U/µL SS II RT; I C.),.µ DTT (.M),.L TP (ATP,GTP,TTP M), µ P CTP ( C/, C/) R (U/, P C., M, WI) C. E b b Q S G- S C (R D C., I, I) C b. DA (GF, GF M; R G, Cb, CA), DA,. T b L Mb (R G), µl P-A (./L, R G), µl C- H DA ( C,./L, I C.) C. T b b b b C. A b, C L X (SSC) b % S (SDS) C.X SSC b % SDS. P-b PI (M D, S, CA) S (M D). S IQ (M D).
6 . Cb T DA b (MCA) (SMC) b (SAH) b (CSF) RA (F. b ). O, b b b b,,. SAH CSF - b MCA SMC. T MCA SMC RA DA. U b. RA T (L T). T RA S b RA. DA b b b. Hb b µ RA. RA.µ/µ C. R X - b (I C., Cb, CA) (U/µ SS II RT; I C.), µ/ L - - (- - P-b, µc ), R (U/, P C., M, WI),.M DTT C. E b b Q S G- S C (R D C., I, I) C. S (GF, GF M; R G, Cb, CA). T b Mb (R G, Cb, CA), µ P-A (./, R G, Cb, C), µ C- H DA ( C,.µ/µ, I C., Cb, CA) C. T b b b b C. A b, C X S (SSC) b % S (SDS) C.X SSC b % SDS. T PI (M D, S, CA) b G GF. O, IQ (M D, S, CA). F TE b C. Hb b, b G PI.. B Pb b b S M Db (://.S.EDU/MA/SMD/). B.. b b b - C/C [B]. M b b (://..//.).
7 . S T. T b b b b b b. H. T H : F, (T). H : I. P-. T - b : S : F b β b -T : (M,.., M ) (T) =,,..., ). S : P (M,.., M ) b b (M τ(),.., M τ() ) -T. D. T B b : β b b =,..., B. S : T - b b b b : = B b=,..,b I(β b > β b ). A. F (),, b. ACKOWLEDGMETS T J K J R. T b ASA CC-. REFERECES. B J.C.,.. S b. P A S. (), - ().. B M.F., L G., S M.B. b:. G R. () - ().. D, A. C., H, D. V. B M A, Cb U P ().. F, J., D, S. A b, # ( ).. S, M., DA M: A P A, O U P ().. Wb, T.G., L, D., B: A P G A G P, S E. A D. B W-I, ().. WEB SITE REFERECES H b. τ,...,.
8 . S M Db ://-.S.EDU/MA/SMD/. B. M b ://-..//.. C B I, E ://.b.../e/ b b b b b b F : T ( b ) TZD-.
9 IDETIFIERS best I: EST :. GB A: AI GB : CLOE IFO C I: IMAGE: ( ) S: IMAGE C, LLL DA : DA SEQUECE TTTTTTTTTTTTTTTACAGGGAAACAGCATTTTTAATGTTTTATTTT TCACTTGTGAAAAATATATATAATATATCTTCCACATACAGAAGAGTC CTGCAGCTTGAGTCAGAGGAAGCTGAAAGAAAGGCACATACAGGGA GCAGATCTTTCCATACAGTTTTCAGTTAAACCAGCATTCAGGGCACA GCAAGTGACAACAAAAGCCCAGGCTGCCTGTGCACACGACTCTGAA GAGAAACATCACAGACAACCCTTAGGTCTACCATGAATGGTTTCTAT TTAATAGACCTATCAGACCACCCGGGAACAAATTGATACCTGAACAA AGACACACCAGGGCAAGACAAAT F : E EST b CBI GB b. T (T). TZD ()..... T (b).... T () T F : () M (T). (T); (b) P, V, b b C. (T),, =,,,..; () b TZD-, b C
10 C. B ().... T (b)... T () T F : T F, b b. Cb ().... T (b)... T () T F : T F, b -.
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