Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel
Detection of double-stranded DNA (dsDNA) and single-strended DNA (ssDNA) and RNA with two new cyanine dyes Cyan 13 and Cyan 40 is reported. Cyan 13 and Cyan. 40 bind to nucleic acids to form a stable fluorescent complexes and can be used for the detection of DMA and RNA samples: separated by gel ele...
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| Date: | 1997 |
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Інститут молекулярної біології і генетики НАН України
1997
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| Cite this: | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel / S.M. Yarmoluk, I.Y. Dubey // Биополимеры и клетка. — 1997. — Т. 13, № 5. — С. 419-421. — Бібліогр.: 7 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1860100453619990528 |
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| author | Yarmoluk, S.M. Dubey, I.Y. |
| author_facet | Yarmoluk, S.M. Dubey, I.Y. |
| citation_txt | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel / S.M. Yarmoluk, I.Y. Dubey // Биополимеры и клетка. — 1997. — Т. 13, № 5. — С. 419-421. — Бібліогр.: 7 назв. — англ. |
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| description | Detection of double-stranded DNA (dsDNA) and single-strended DNA (ssDNA) and RNA with two new cyanine dyes Cyan 13 and Cyan 40 is reported. Cyan 13 and Cyan. 40 bind to nucleic acids to form a stable fluorescent complexes and can be used for the detection of DMA and RNA samples: separated by gel electrophoresis. Sensitivity of detection is comparable to tfat for ethidium bromide (EtBr), a common nucleic add staining dye.
Два нових ціанінових барвники Cyan 13 та Cyan 40 було застосовано для флюоресцентної детекцїі двоспіральної ДНК та односпіральних ДНК і РНК в електрофорезних гелях. Чутливість детекції нуклеїнових кислот цими барвниками схожа з такою бромистого етидію, що звичайно використовується для цієї мети.
Два новых цианиновых красителя Cyan 13 и Cyan 40 применены для флюоресцентной детекции двуспиральной ДНК и одно- спиральных ДНК и РНК в электрофорезных гелях. Чувствительность детекции нуклеиновых кислот этими красителями подобна таковой бромистого этидия, который обычно используется для этой цели.
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ISSN 0233-7657. Биополимеры и клетка. 1997. Т. 13. № 5
МЕТОДЫ
Interaction of cyanine dyes with nucleic acids.
3, The use of new cyanine dyes Cyan 13 and
Cyan 40 for detection of nucleic acids in agarose gel
Sergiy M. Yarmoluk, Igor Ya. Dubey 1
Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine
150 Zabolotnogo str., Kyiv, 252143 , Ukraine
1
Institute of Bioorganic Chemistry and Petrochemistry. National Academy of Sciences of Ukraine
1 Murmanska str., Kyiv, 253660, Ukraine
Detection of double-stranded DNA (dsDNA) and single-strcnded DNA (ssDNA) and UNA with two new
cyanine dyes Cyan 13 and Cyan 40 is reported. Cyan 13 and Cyan 40 bind to nucleic acids to form a
stable fluorescent complexes and can be used for the detection of DMA and RNA samples separated by gel
electrophoresis. Sensitivity of detection is comparable to tl at for ethidium bromide (EtBr), a common
nucleic acid staining dye.
Materials and Methods DNA and RNA preparation.
Double-stranded plasmid DNAs pBR322 and pUC18
were isolated by standard alkaline lysis method of
Birnboim [I ]. Linear plasmid DNA pUCJS was ob
tained by digestion with BamHI restrietase. Closed
circular single-stranded M13mpl DNA (M13 ssDNA)
was obtained from USB (USA). Total RNA was
extracted from rat liver tissue by guanidinium thio-
cyanate-phenol method according to Chomczynski et
al. [2].
Agarose gel electrophoresis and staining of nucleic
acids. 0.8 % Agarose gels were prepared in standard
TAE buffer (40 raM Tris-acetate, 1 mM EDTA, pH
8.0) [3]. DNA and RNA samples in TAE buffer were
loaded on the gel after adding tracking dyes xylene
cyanole and bromophenole blue in 30 % glycerol to
concentration 0,025 %. Separations were carried out
at 10 V/cm. DNA and RNA gels were stained after
electrophoresis in the solution of Cyan 13, Cyan 40
or EtBr (1 fig/m\) for 30 min.
Fluorescence detection of DNA and RNA in
agarose gels. Detection of DNA-dye and RNA-dye
complexes was performed with UV-transilluminator
2011 Macrovue («LKB», Sweden) at 524 nm. The gels
were photographed using orange filter.
© S. M. YARMOLUK, f. Ya. D U B E Y , 1997
Results and Discussion. The use of non-isotopic
detection techniques in biology and medicine is an
attractive alternative to radioactive detection methods.
Cyanine dyes ars now widely used as fluorescence
detection probes. Monomethyne cyanine dyes are
weakly fluorescent in ree state but show strong
fluorescence upon binding to DNA or RNA, and some
of these reagents are the most sensitive nucleic acid
stains currently available [4, 5 ] . A series of new
fluorescent cyanine dyes with improved spectroscopic
properties including Cyan 13 [6] and Cyan 40 [7]
has been recently developed in our laboratory (Fig.
1).
In this communication we demonstrate that Cyan
13 and Cyan 40 can be successfully used for DNA
and RNA detection in the standard procedures. Dyes
bind efficiently to different forms of DNA — co-
valently closed circular and linear dsDNA and
ssDNA, as well as to various RNA types — ribosomal
RNA, mRNA and tRNA, forming stable fluorescent
complexes.
In Fig. 2 is shown the comparison of DNA
staining in agarose gel with Cyan 13, Cyan 40 and
EtBr. It was shown that fluorescence of Cyan 13 and
Cyan 40 complexes with linear dsDNA and circular
M13 ssDNA (lanes 1 and J, respectively) was com
parable wit that of EtBr.
419
YARMOLUK. S. M., D U B B Y I. Yet.
I
Fig. 2. Detection of DNA in agarose gels after electrophoresis:, lane
/ — 200 ng of dsDNA pUC18 linearized with BaniHI restrictase;
lane 2 — 300 ng of intact pBR322 dsDNA; lane 3 — 200 ng of M13
ssDNA. Staining was made with Cyan 13 (A), with Cyan 40 (B) and
EtBr (C)
But the intensity of fluorescence of EtBr complex
with circular closed dsDNA (lane 2) was higher than
those for Cyan 13 and Cyan 40. It is necessary to
note that the contamination of bacterial tRNAs in
samples of circular closed plasmid DNAs shined with
almost equal intensity.
The fluorescence intensity of Cyan 13 and Cyan
40 complexes with RNA is enough high. As is shown
in Fig. 3, all types of RNA — ribosomal RNA, mRNA
and tRNA — can be detected in total RNA samples.
Both 28S and 18S rRNAs were detectable in the gels
stained with Cyan 13 and Cyan 40, as well as stained
with EtBr. The fluorescence of RNA complexes with
Cyan 40 was lower as compared with two other dyes.
The colours of fluorescence of RNA and DNA
N
А В С
Fig. 3. Staining of RNA in agarose gels after electrophoresis with
Cyan 13 (Л), Cyan 40 (B) and EtBr (C) . Each lane contains 5 ,wg
of total rat liver RNA. Positions of ribosomal 28S and 18S RNA are
indicated
complexes with Cyan 13 and Cyan 40 were from blue
to light green. The colour of Cyan 13 — DNA com
plexes was shifted to red by increasing dye con
centration or time of staining. It is possible that such
a change off luorescence spectrum was caused by
formation of H-aggregates (unpublished results).
It can be concluded therefore that two new
cyanine dyes Cyan 13 and Cyan 40 can be used for
DNA and RNA detection in agarose gels.
420
INTERACTION O F C Y A N I N E D Y E S WITH NUCLEIC ACIDS
Acknowledgements. We are indebted to Dr.
Yu. Kovtun for providing the dyes and thank Ms.
V. Koval'ska for technical assistance.
С M. Ярмолюк, І. Я. Дубей
Взаємодія ціанінових барвників з нуклеїновими кислотами.
3. Застосування нових ціанінових барвників Cyan 13 та Cyan 40
для визначення нуклеїнових кислот в агарозних гелях
Резюме
Два нових ціанінових барвники Cyan 13 та Cyan 40 було
застосовано для флюоресцентно)' детекцїі двоспіральної ДНК
та односпіральних ДНК і РНК в електрофорезних гелях.
Чутливість детекції нуклеїнових кислот цими барвниками
схожа з такою бромистого етидію, що звичайно використо
вується для цієї мети.
С. Н. Ярмолюк, И. Я. Дубей
Взаимодействие цианиновых красителей с нуклеиновыми
кислотами. 3. Применение новых цианиновых красителей
Cyan 13 и Cyan 40 для определения нуклеиновых кислот
в агарозных гелях
Резюме
Два новых цианиновых красителя Cyan 13 и Cyan 40 примене
ны для флюоресцентной детекции двуспиральной ДНК и одно-
спиральных ДНК и РНК в электрофорезных гелях. Чувстви
тельность детекции нуклеиновых кислот этими красителями
подобна таковой бромистого этидия, который обычно исполь
зуется для этой цели.
REFERENCES
1 Birnboim Н. A rapid alkaline extraction method for the
isolation of plasraid DNA / / Meth. Enzymol .—1983.—100 —
P. 243—256 .
2. Chomczynski P., Sacchi N. Single-step method of RNA
isolation by acid guanidinium thiocyanate — phenol — chloro
form extraction / / Anal. B iochem.—1987 .—162 .—P. 156—
159.
3. Maniatis Т., Fritsch E. F., Sambrook J. Molecular cloning: A
laboratory manual.—New York: Cold Spring Harbor Lab.,
1982 .—545 p.
4. Haughland R. P. Molecular probes. Handbook of fluorescent
probes and research chemicals.—Eugene, OR., 1996 .—679 p.
5. Lee G. L., Chen C.-H., Chiu L. A. Thiazole Orange: a new
dye for reticulocyte analysis / / Cytometry.—1986.—7.—
P. 5 0 8 — 5 1 7 .
6. Yarmoluk S. M., Zhyvoloup A. N., Kovatska V. B. et at
Interaction of cyanine dyes with nucleic acids. I. Studies on
monomethyne cyanine dyes as possible fluorescent probes for
the detection of nucleic acids / / Biopolymers and Cell.—
1996 .—12, N 1.—P. 6 9 — 7 4 .
7. Yarmoluk S. M., Kovalska V. В., Smirnova Т. V. et at.
Interaction of cyanine dyes with nucleic acids. II. Spectroscopic
properties of methyleneoxy analogues of Thiazole Orange / /
Ibid.—N 6.—P. 7 4 — 8 1 .
Received 18.02.97
421
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| id | nasplib_isofts_kiev_ua-123456789-155646 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 0233-7657 |
| language | English |
| last_indexed | 2025-12-07T17:28:10Z |
| publishDate | 1997 |
| publisher | Інститут молекулярної біології і генетики НАН України |
| record_format | dspace |
| spelling | Yarmoluk, S.M. Dubey, I.Y. 2019-06-17T09:41:40Z 2019-06-17T09:41:40Z 1997 Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel / S.M. Yarmoluk, I.Y. Dubey // Биополимеры и клетка. — 1997. — Т. 13, № 5. — С. 419-421. — Бібліогр.: 7 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0004A1 https://nasplib.isofts.kiev.ua/handle/123456789/155646 Detection of double-stranded DNA (dsDNA) and single-strended DNA (ssDNA) and RNA with two new cyanine dyes Cyan 13 and Cyan 40 is reported. Cyan 13 and Cyan. 40 bind to nucleic acids to form a stable fluorescent complexes and can be used for the detection of DMA and RNA samples: separated by gel electrophoresis. Sensitivity of detection is comparable to tfat for ethidium bromide (EtBr), a common nucleic add staining dye. Два нових ціанінових барвники Cyan 13 та Cyan 40 було застосовано для флюоресцентної детекцїі двоспіральної ДНК та односпіральних ДНК і РНК в електрофорезних гелях. Чутливість детекції нуклеїнових кислот цими барвниками схожа з такою бромистого етидію, що звичайно використовується для цієї мети. Два новых цианиновых красителя Cyan 13 и Cyan 40 применены для флюоресцентной детекции двуспиральной ДНК и одно- спиральных ДНК и РНК в электрофорезных гелях. Чувствительность детекции нуклеиновых кислот этими красителями подобна таковой бромистого этидия, который обычно используется для этой цели. We are indebted to Dr. Yu. Kovtun for providing the dyes and thank Ms. V. Koval'ska for technical assistance. en Інститут молекулярної біології і генетики НАН України Биополимеры и клетка Методы Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel Взаємодія ціанінових барвників з нуклеїновими кислотами. 3. Застосування нових ціанінових барвників Cyan 13 та Cyan 40 для визначення нуклеїнових кислот в агарозних гелях Взаимодействие цианиновых красителей с нуклеиновыми кислотами. 3. Применение новых цианиновых красителей Cyan 13 и Cyan 40 для определения нуклеиновых кислот в агарозных гелях Article published earlier |
| spellingShingle | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel Yarmoluk, S.M. Dubey, I.Y. Методы |
| title | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel |
| title_alt | Взаємодія ціанінових барвників з нуклеїновими кислотами. 3. Застосування нових ціанінових барвників Cyan 13 та Cyan 40 для визначення нуклеїнових кислот в агарозних гелях Взаимодействие цианиновых красителей с нуклеиновыми кислотами. 3. Применение новых цианиновых красителей Cyan 13 и Cyan 40 для определения нуклеиновых кислот в агарозных гелях |
| title_full | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel |
| title_fullStr | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel |
| title_full_unstemmed | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel |
| title_short | Interaction of cyanine dyes with nucleic acids. 3. The of new cyanine dyes Cyan 13 and Cyan 40 for detection of nucleic acids in agarose gel |
| title_sort | interaction of cyanine dyes with nucleic acids. 3. the of new cyanine dyes cyan 13 and cyan 40 for detection of nucleic acids in agarose gel |
| topic | Методы |
| topic_facet | Методы |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/155646 |
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