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Lina De Smet, Fani Hatjina, Pavlos Ioannidis, Anna Hamamtzoglou, Karel Schoonvaere, Frédéric Francis, Ivan Meeus, Guy Smagghe, Dirk C. de Graaf
Stress indicator gene expression profiles, colony dynamics and tissue development of honey bees exposed to sub-lethal doses of imidacloprid in laboratory and field experiments
www.plosone.org
www.plosone.org
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(17)Tj
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()Tj
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1.627 0 Td
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2.3357 0 Td
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(\(HPLC-E-)Tj
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1.8709 0 Td
[()]TJ
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(the)Tj
1.4683 0 Td
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(1.35)Tj
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1.8708 0 Td
[()]TJ
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(g/kg.)Tj
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5.4651 0 Td
(was)Tj
1.7008 0 Td
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2.6192 0 Td
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1.0375 0 Td
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1.0148 0 Td
(3.1)Tj
1.3947 0 Td
(and)Tj
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(206.7)Tj
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2.3527 0 Td
[()]TJ
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1.0318 0 Td
(pollen)Tj
2.7099 0 Td
(patty)Tj
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(and)Tj
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3.0103 0 Td
(with)Tj
1.9956 0 Td
(5x)Tj
1.1622 0 Td
(SSC)Tj
1.8369 0 Td
(\(0.15)Tj
2.211 0 Td
(M)Tj
1.1111 0 Td
(NaCl)Tj
2.8177 0 Td
(0.015)Tj
2.347 0 Td
(M)Tj
1.1112 0 Td
(sodium)Tj
3.2315 0 Td
(citrate)Tj
2.7382 0 Td
(for)Tj
1.3833 0 Td
(1x)Tj
1.1622 0 Td
(SSC\))Tj
2.177 0 Td
(and)Tj
-34.9112 -1.2982 Td
(0.1%)Tj
2.1543 0 Td
(sodium)Tj
3.2258 0 Td
(dodecyl)Tj
3.3335 0 Td
(sulphate,)Tj
3.7814 0 Td
(the)Tj
1.4683 0 Td
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1.0374 0 Td
(the)Tj
1.4684 0 Td
(cassette.)Tj
3.4696 0 Td
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(cDNA)Tj
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(corre-)Tj
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3.9628 0 Td
(with)Tj
1.9955 0 Td
(3)Tj
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0.6974 0 Td
[()]TJ
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(g)Tj
1.1905 0 Td
(total)Tj
2.0012 0 Td
(RNA)Tj
2.2621 0 Td
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1.7008 0 Td
(mixed)Tj
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(hybridization)Tj
5.6069 0 Td
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(\(final)Tj
2.3414 0 Td
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(25%)Tj
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4.9039 0 Td
(5x)Tj
1.1565 0 Td
(SSC)Tj
1.8368 0 Td
(and)Tj
1.7291 0 Td
(0.1%)Tj
2.1544 0 Td
(SDS\))Tj
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1.0204 0 Td
(a)Tj
0.6577 0 Td
(final)Tj
2.0012 0 Td
(volume)Tj
3.1975 0 Td
(of)Tj
1.0148 0 Td
(75)Tj
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1.1735 0 Td
[()]TJ
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(and)Tj
1.7291 0 Td
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1.7008 0 Td
(heated)Tj
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1.389 0 Td
(1)Tj
0.6917 0 Td
(minute)Tj
3.1067 0 Td
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(65C)Tj
2.228 0 Td
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1.0204 0 Td
(denature)Tj
3.7758 0 Td
(the)Tj
1.4683 0 Td
(cDNA)Tj
2.8006 0 Td
(and)Tj
1.7348 0 Td
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(cooled)Tj
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1.2755 0 Td
(ice)Tj
1.3323 0 Td
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1.389 0 Td
(30)Tj
1.1735 0 Td
(sec.)Tj
1.6498 0 Td
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1.7858 0 Td
(labelled)Tj
3.2711 0 Td
(cDNA)Tj
2.8007 0 Td
(was)Tj
1.7007 0 Td
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1.0261 0 Td
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1.4684 0 Td
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2.2847 0 Td
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(overnight)Tj
4.0875 0 Td
(at)Tj
0.9524 0 Td
(33.5C.)Tj
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5.8337 0 Td
(the)Tj
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3.1804 0 Td
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2.3131 0 Td
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0.9524 0 Td
(45C)Tj
2.228 0 Td
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(5)Tj
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(2x)Tj
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(and)Tj
1.7291 0 Td
(0.1%)Tj
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(SDS,)Tj
2.1373 0 Td
(twice)Tj
2.3131 0 Td
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1.9956 0 Td
(0.5x)Tj
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(and)Tj
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0.6973 0 Td
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0.9524 0 Td
(room)Tj
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1.9956 0 Td
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5.142 0 Td
(detection)Tj
3.9401 0 Td
(is)Tj
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(a)Tj
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(three-step)Tj
4.2122 0 Td
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1.1055 0 Td
(the)Tj
1.474 0 Td
(first)Tj
1.8029 0 Td
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2.126 0 Td
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(1%)Tj
1.4457 0 Td
(blocking)Tj
3.6793 0 Td
(solution)Tj
3.481 0 Td
(\(Roche\))Tj
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1.3833 0 Td
(30)Tj
1.1679 0 Td
(min)Tj
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(to)Tj
1.0205 0 Td
(prevent)Tj
3.2541 0 Td
(nonspecific)Tj
4.7906 0 Td
(attraction)Tj
4.0988 0 Td
(of)Tj
1.0092 0 Td
(the)Tj
1.4683 0 Td
(antibody)Tj
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(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(membrane.)Tj
4.7905 0 Td
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1.1112 0 Td
(the)Tj
1.4684 0 Td
(following)Tj
3.9798 0 Td
(step)Tj
1.8198 0 Td
(the)Tj
1.4683 0 Td
(membranes)Tj
4.9266 0 Td
(were)Tj
2.126 0 Td
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4.1782 0 Td
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1.9956 0 Td
(1500x)Tj
2.5852 0 Td
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-32.944 -1.2982 Td
(anti-digoxigenin)Tj
6.8711 0 Td
(\(Roche\))Tj
3.4015 0 Td
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1.0375 0 Td
(1%)Tj
1.4513 0 Td
(blocking)Tj
3.6794 0 Td
(solution.)Tj
3.7077 0 Td
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1.7858 0 Td
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4.3597 0 Td
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4.6317 0 Td
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1.0262 0 Td
(alkaline)Tj
-34.0722 -1.2983 Td
(phosphatase)Tj
5.125 0 Td
(which)Tj
2.6532 0 Td
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2.7552 0 Td
(colorimetric)Tj
5.142 0 Td
(development)Tj
5.4142 0 Td
(possible.)Tj
3.634 0 Td
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1.7801 0 Td
(unbound)Tj
3.9175 0 Td
(antibody)Tj
3.7644 0 Td
(was)Tj
-34.1856 -1.3039 Td
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3.1747 0 Td
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2.2281 0 Td
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1.0374 0 Td
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1.7065 0 Td
(washing)Tj
3.5092 0 Td
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2.1827 0 Td
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1.9956 0 Td
(PBS)Tj
1.8368 0 Td
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1.3833 0 Td
(15)Tj
1.1736 0 Td
(min)Tj
1.8538 0 Td
(after)Tj
2.0353 0 Td
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2.6475 0 Td
(the)Tj
1.474 0 Td
(slides)Tj
2.4038 0 Td
(were)Tj
2.1259 0 Td
(equilibrated)Tj
-32.7682 -1.2983 Td
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1.0318 0 Td
(TBS)Tj
1.8878 0 Td
(buffer)Tj
2.6249 0 Td
(for)Tj
1.3889 0 Td
(5)Tj
0.6974 0 Td
(min.)Tj
2.0806 0 Td
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1.1055 0 Td
(the)Tj
1.4683 0 Td
(last)Tj
1.5591 0 Td
(step,)Tj
2.0466 0 Td
(the)Tj
1.4683 0 Td
(membrane)Tj
4.5694 0 Td
(carrying)Tj
3.5547 0 Td
(the)Tj
1.4683 0 Td
(hybridized)Tj
4.507 0 Td
(probe)Tj
2.5512 0 Td
(and)Tj
-34.0098 -1.2983 Td
(bound)Tj
2.8346 0 Td
(antibody)Tj
3.7644 0 Td
(conjugate)Tj
4.1045 0 Td
(was)Tj
1.7008 0 Td
(reacted)Tj
3.1237 0 Td
(with)Tj
1.9956 0 Td
(the)Tj
1.4684 0 Td
(colorimetric)Tj
5.1476 0 Td
(detection)Tj
3.9345 0 Td
(reagents)Tj
3.549 0 Td
(NBT)Tj
2.1429 0 Td
(\(nitro)Tj
-0.019 Tc
-33.766 -1.3039 Td
(blue)Tj
1.8651 0 Td
(tetrazolium)Tj
4.6602 0 Td
(salt\))Tj
1.8255 0 Td
(and)Tj
1.6837 0 Td
(BCIP)Tj
2.3018 0 Td
(\(5-bromo-4-chloro-3-indo)Tj
10.3067 0 Td
(lyl)Tj
1.1225 0 Td
(phosphate\).)Tj
4.7338 0 Td
(The)Tj
1.7348 0 Td
(NBT/BCIP)Tj
4.5014 0 Td
(stock)Tj
-34.7355 -1.2982 Td
(solution)Tj
3.3618 0 Td
(\(Roche\))Tj
3.2995 0 Td
(contained)Tj
4.0592 0 Td
(18.75)Tj
2.2734 0 Td
(mg/ml)Tj
2.8289 0 Td
(nitro)Tj
2.1374 0 Td
(blue)Tj
1.8651 0 Td
(tetrazolium)Tj
4.6602 0 Td
(chloride)Tj
3.3958 0 Td
(and)Tj
1.6838 0 Td
(9.4)Tj
1.3493 0 Td
(mg/ml)Tj
2.829 0 Td
(5-bromo-)Tj
-33.7434 -1.2983 Td
(4-chloro-3-indolyl-phosph)Tj
10.318 0 Td
(ate,)Tj
1.5364 0 Td
(toluidine-salt)Tj
5.3177 0 Td
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1.0035 0 Td
(67%)Tj
1.8822 0 Td
(DMSO)Tj
3.0104 0 Td
(\(v/v\).)Tj
2.279 0 Td
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1.7348 0 Td
(slides)Tj
2.3187 0 Td
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2.0579 0 Td
(incubated)Tj
4.0422 0 Td
(in)Tj
1.0092 0 Td
(15)Tj
-36.51 -1.3039 Td
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1.2529 0 Td
(TBS)Tj
1.8425 0 Td
(buffer)Tj
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(300)Tj
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1.5987 0 Td
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(l)Tj
0.9468 0 Td
(NBT/BCIP)Tj
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(for)Tj
1.3436 0 Td
(30)Tj
1.1395 0 Td
(min.)Tj
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(18.75)Tj
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(nitro)Tj
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(blue)Tj
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(tetra-)Tj
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(zolium)Tj
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(chloride)Tj
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(and)Tj
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(9.4)Tj
1.355 0 Td
(mg/ml)Tj
2.8233 0 Td
(5-bromo-4-chloro-3-indoly)Tj
10.6525 0 Td
(l-phosphate,)Tj
4.9719 0 Td
(toluidine-salt)Tj
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(in)Tj
1.0092 0 Td
(67%)Tj
-34.1119 -1.2983 Td
(\(DMSO\))Tj
3.6623 0 Td
(\(v/v\).)Tj
2.2734 0 Td
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3.4752 0 Td
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(18.75)Tj
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(nitro)Tj
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(blue)Tj
1.8652 0 Td
(tetrazolium)Tj
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(chloride)Tj
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(and)Tj
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(9.4)Tj
1.355 0 Td
(mg/ml)Tj
2.8232 0 Td
(5-bromo-)Tj
-33.4145 -1.2982 Td
(4-chloro-3-indolyl-phosph)Tj
10.318 0 Td
(ate,)Tj
1.5364 0 Td
(toluidine-salt)Tj
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1.0035 0 Td
(67%)Tj
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(\(DMSO\))Tj
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(\(v/v\).)Tj
2.2734 0 Td
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(mg/ml)Tj
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(nitro)Tj
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(blue)Tj
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(tetrazolium)Tj
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(chloride)Tj
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(and)Tj
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(9.4)Tj
1.3493 0 Td
(mg/ml)Tj
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(5-bromo-4-chloro-3-indolyl-phosp)Tj
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(hate,)Tj
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(toluidine-salt)Tj
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(in)Tj
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(67%)Tj
1.8821 0 Td
(\(DMSO\))Tj
3.6624 0 Td
(\(v/v\).)Tj
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2.2733 0 Td
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(5-)Tj
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(bromo-4-chloro-3-indolyl-p)Tj
10.9246 0 Td
(hosphate,)Tj
3.9061 0 Td
(toluidine-salt)Tj
5.3121 0 Td
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1.0035 0 Td
(67%)Tj
1.8822 0 Td
(\(DMSO\))Tj
3.6623 0 Td
(\(v/v\).The)Tj
3.7927 0 Td
(reaction)Tj
3.3732 0 Td
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3.2654 0 Td
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1.1509 0 Td
(washing)Tj
3.4016 0 Td
(the)Tj
1.4286 0 Td
(slides)Tj
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1.0091 0 Td
(distilled)Tj
3.2485 0 Td
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(centrifugation.)Tj
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(quently,)Tj
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(the)Tj
1.4287 0 Td
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(scanned)Tj
3.3846 0 Td
(using)Tj
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(the)Tj
1.423 0 Td
(ArrayIT)Tj
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(1)Tj
/F7 1 Tf
4.3653 0 Td
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 444.9259 295.8236 cm
BT
/F7 1 Tf
1 TL
-0.0253 Tc
0 0 Td
(TM)Tj
ET
Q
q
1 j
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BT
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(microarray)Tj
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Q
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(inverted)Tj
3.4299 0 Td
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(images)Tj
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(\(Innopsys\))Tj
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(ascribe)Tj
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(value)Tj
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(the)Tj
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(data)Tj
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(normalized)Tj
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(and)Tj
1.6951 0 Td
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(arrays)Tj
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(package)Tj
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(control)Tj
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