Microfluidics-based in situ Padlock/Rolling Circle Amplification System for Counting Single DNA Molecules in a Cell
| العنوان: | Microfluidics-based in situ Padlock/Rolling Circle Amplification System for Counting Single DNA Molecules in a Cell |
|---|---|
| المؤلفون: | Yuri Ishigaki, Kiichi Sato, Mats Nilsson, Arisa Kuroda, Kae Sato |
| المصدر: | Analytical Sciences. 30:1107-1112 |
| بيانات النشر: | Springer Science and Business Media LLC, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | In situ, Cells, Microfluidics, Nanotechnology, DNA, Nucleic acid amplification technique, Analytical Chemistry, chemistry.chemical_compound, chemistry, Rolling circle replication, Humans, Molecule, Biological system, Nucleic Acid Amplification Techniques, HeLa Cells |
| الوصف: | In situ padlock/rolling circle amplification (RCA) is a method used to amplify, visualize, and quantify target DNA molecules in cells. However, the multiple reaction steps involved make this technique costly and cumbersome. We developed a novel, simplified, automated microfluidic system for RCA, and demonstrated its effectiveness by counting amplified mitochondrial DNA fragments in HeLa cells. After optimizing the volume of the reaction solutions and washing buffer composition, the product yield was equal to that obtained by the conventional manual method. The required volume of reagents was reduced to 10 μL, which is less than half the volume used in the conventional method. To the best of our knowledge, this is the first report of an automated microfluidic method for in situ padlock/RCA, which can be useful for making highly efficient pathological diagnoses. |
| تدمد: | 1348-2246 0910-6340 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::417e150a75c41fa8359e534d8eca2e4dTest https://doi.org/10.2116/analsci.30.1107Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....417e150a75c41fa8359e534d8eca2e4d |
| قاعدة البيانات: | OpenAIRE |
| ResultId |
1 |
|---|---|
| Header |
edsair OpenAIRE edsair.doi.dedup.....417e150a75c41fa8359e534d8eca2e4d 813 3 unknown 812.963806152344 |
| PLink |
https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&scope=site&db=edsair&AN=edsair.doi.dedup.....417e150a75c41fa8359e534d8eca2e4d&custid=s6537998&authtype=sso |
| FullText |
Array
(
[Availability] => 0
)
|
| Items |
Array
(
[Name] => Title
[Label] => Title
[Group] => Ti
[Data] => Microfluidics-based in situ Padlock/Rolling Circle Amplification System for Counting Single DNA Molecules in a Cell
)
Array ( [Name] => Author [Label] => Authors [Group] => Au [Data] => <searchLink fieldCode="AR" term="%22Yuri+Ishigaki%22">Yuri Ishigaki</searchLink><br /><searchLink fieldCode="AR" term="%22Kiichi+Sato%22">Kiichi Sato</searchLink><br /><searchLink fieldCode="AR" term="%22Mats+Nilsson%22">Mats Nilsson</searchLink><br /><searchLink fieldCode="AR" term="%22Arisa+Kuroda%22">Arisa Kuroda</searchLink><br /><searchLink fieldCode="AR" term="%22Kae+Sato%22">Kae Sato</searchLink> ) Array ( [Name] => TitleSource [Label] => Source [Group] => Src [Data] => <i>Analytical Sciences</i>. 30:1107-1112 ) Array ( [Name] => Publisher [Label] => Publisher Information [Group] => PubInfo [Data] => Springer Science and Business Media LLC, 2014. ) Array ( [Name] => DatePubCY [Label] => Publication Year [Group] => Date [Data] => 2014 ) Array ( [Name] => Subject [Label] => Subject Terms [Group] => Su [Data] => <searchLink fieldCode="DE" term="%22In+situ%22">In situ</searchLink><br /><searchLink fieldCode="DE" term="%22Cells%22">Cells</searchLink><br /><searchLink fieldCode="DE" term="%22Microfluidics%22">Microfluidics</searchLink><br /><searchLink fieldCode="DE" term="%22Nanotechnology%22">Nanotechnology</searchLink><br /><searchLink fieldCode="DE" term="%22DNA%22">DNA</searchLink><br /><searchLink fieldCode="DE" term="%22Nucleic+acid+amplification+technique%22">Nucleic acid amplification technique</searchLink><br /><searchLink fieldCode="DE" term="%22Analytical+Chemistry%22">Analytical Chemistry</searchLink><br /><searchLink fieldCode="DE" term="%22chemistry%2Echemical%5Fcompound%22">chemistry.chemical_compound</searchLink><br /><searchLink fieldCode="DE" term="%22chemistry%22">chemistry</searchLink><br /><searchLink fieldCode="DE" term="%22Rolling+circle+replication%22">Rolling circle replication</searchLink><br /><searchLink fieldCode="DE" term="%22Humans%22">Humans</searchLink><br /><searchLink fieldCode="DE" term="%22Molecule%22">Molecule</searchLink><br /><searchLink fieldCode="DE" term="%22Biological+system%22">Biological system</searchLink><br /><searchLink fieldCode="DE" term="%22Nucleic+Acid+Amplification+Techniques%22">Nucleic Acid Amplification Techniques</searchLink><br /><searchLink fieldCode="DE" term="%22HeLa+Cells%22">HeLa Cells</searchLink> ) Array ( [Name] => Abstract [Label] => Description [Group] => Ab [Data] => In situ padlock/rolling circle amplification (RCA) is a method used to amplify, visualize, and quantify target DNA molecules in cells. However, the multiple reaction steps involved make this technique costly and cumbersome. We developed a novel, simplified, automated microfluidic system for RCA, and demonstrated its effectiveness by counting amplified mitochondrial DNA fragments in HeLa cells. After optimizing the volume of the reaction solutions and washing buffer composition, the product yield was equal to that obtained by the conventional manual method. The required volume of reagents was reduced to 10 μL, which is less than half the volume used in the conventional method. To the best of our knowledge, this is the first report of an automated microfluidic method for in situ padlock/RCA, which can be useful for making highly efficient pathological diagnoses. ) Array ( [Name] => ISSN [Label] => ISSN [Group] => ISSN [Data] => 1348-2246<br />0910-6340 ) Array ( [Name] => URL [Label] => Access URL [Group] => URL [Data] => <link linkTarget="URL" linkTerm="https://explore.openaire.eu/search/publication?articleId=doi_dedup___::417e150a75c41fa8359e534d8eca2e4d" linkWindow="_blank">https://explore.openaire.eu/search/publication?articleId=doi_dedup___::417e150a75c41fa8359e534d8eca2e4d</link><br /><link linkTarget="URL" linkTerm="https://doi.org/10.2116/analsci.30.1107" linkWindow="_blank">https://doi.org/10.2116/analsci.30.1107</link> ) Array ( [Name] => Copyright [Label] => Rights [Group] => Cpyrght [Data] => OPEN ) Array ( [Name] => AN [Label] => Accession Number [Group] => ID [Data] => edsair.doi.dedup.....417e150a75c41fa8359e534d8eca2e4d ) |
| RecordInfo |
Array
(
[BibEntity] => Array
(
[Languages] => Array
(
[0] => Array
(
[Text] => Undetermined
)
)
[PhysicalDescription] => Array
(
[Pagination] => Array
(
[PageCount] => 6
[StartPage] => 1107
)
)
[Subjects] => Array
(
[0] => Array
(
[SubjectFull] => In situ
[Type] => general
)
[1] => Array
(
[SubjectFull] => Cells
[Type] => general
)
[2] => Array
(
[SubjectFull] => Microfluidics
[Type] => general
)
[3] => Array
(
[SubjectFull] => Nanotechnology
[Type] => general
)
[4] => Array
(
[SubjectFull] => DNA
[Type] => general
)
[5] => Array
(
[SubjectFull] => Nucleic acid amplification technique
[Type] => general
)
[6] => Array
(
[SubjectFull] => Analytical Chemistry
[Type] => general
)
[7] => Array
(
[SubjectFull] => chemistry.chemical_compound
[Type] => general
)
[8] => Array
(
[SubjectFull] => chemistry
[Type] => general
)
[9] => Array
(
[SubjectFull] => Rolling circle replication
[Type] => general
)
[10] => Array
(
[SubjectFull] => Humans
[Type] => general
)
[11] => Array
(
[SubjectFull] => Molecule
[Type] => general
)
[12] => Array
(
[SubjectFull] => Biological system
[Type] => general
)
[13] => Array
(
[SubjectFull] => Nucleic Acid Amplification Techniques
[Type] => general
)
[14] => Array
(
[SubjectFull] => HeLa Cells
[Type] => general
)
)
[Titles] => Array
(
[0] => Array
(
[TitleFull] => Microfluidics-based in situ Padlock/Rolling Circle Amplification System for Counting Single DNA Molecules in a Cell
[Type] => main
)
)
)
[BibRelationships] => Array
(
[HasContributorRelationships] => Array
(
[0] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Yuri Ishigaki
)
)
)
[1] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Kiichi Sato
)
)
)
[2] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Mats Nilsson
)
)
)
[3] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Arisa Kuroda
)
)
)
[4] => Array
(
[PersonEntity] => Array
(
[Name] => Array
(
[NameFull] => Kae Sato
)
)
)
)
[IsPartOfRelationships] => Array
(
[0] => Array
(
[BibEntity] => Array
(
[Dates] => Array
(
[0] => Array
(
[D] => 01
[M] => 12
[Type] => published
[Y] => 2014
)
)
[Identifiers] => Array
(
[0] => Array
(
[Type] => issn-print
[Value] => 13482246
)
[1] => Array
(
[Type] => issn-print
[Value] => 09106340
)
[2] => Array
(
[Type] => issn-locals
[Value] => edsair
)
[3] => Array
(
[Type] => issn-locals
[Value] => edsairFT
)
)
[Numbering] => Array
(
[0] => Array
(
[Type] => volume
[Value] => 30
)
)
[Titles] => Array
(
[0] => Array
(
[TitleFull] => Analytical Sciences
[Type] => main
)
)
)
)
)
)
)
|
| IllustrationInfo |