New quick method for isolating RNA from laser captured cells stained by immunofluorescent immunohistochemistry; RNA suitable for direct use in fluorogenic TaqMan one-step real-time RT-PCR

dc.contributor.author Gallup, Jack
dc.contributor.author Gallup, Jack
dc.contributor.author Kawashima, Kenji
dc.contributor.author Lucero, Ginger
dc.contributor.author Ackermann, Mark
dc.contributor.author Ackermann, Mark
dc.contributor.department Veterinary Pathology
dc.date 2018-02-13T06:19:44.000
dc.date.accessioned 2020-07-07T05:16:09Z
dc.date.available 2020-07-07T05:16:09Z
dc.date.copyright Sat Jan 01 00:00:00 UTC 2005
dc.date.embargo 2013-02-25
dc.date.issued 2005-01-01
dc.description.abstract <p>We describe a new approach for reliably isolating one-step real-time quantitative RT-PCR-quality RNA from laser captured cells retrieved from frozen sections previously subjected to immunofluorescent immunohistochemistry (IF-IHC) and subsequently subjected to fluorogenic one-step real-time RT-PCR analysis without the need for costly, time-consuming linear amplification. One cell’s worth of RNA can now be interrogated with confidence. This approach represents an amalgam of technologies already offered commercially by Applied Biosystems, Arcturus and Invitrogen. It is the primary focus of this communication to expose the details and execution of an important new LCM RNA isolation technique, but also provide a detailed account of the IF-IHC procedure preceding RNA isolation, and provide information regarding our approach to fluorogenic one-step real-time RT-PCR in general. Experimental results shown here are meant to supplement the primary aim and are not intended to represent a complete scientific study. It is important to mention, that since LCM-RT-PCR is still far less expensive than micro-array analysis, we feel this approach to isolating RNA from LCM samples will be of continuing use to many researchers with limited budgets in the years ahead.</p>
dc.description.comments <p>This article is from <em>Biological Procedures Online </em>7, no. 1 (2005): 70–92, doi:<a href="http://dx.doi.org/10.1251/bpo107" target="_blank">10.1251/bpo107</a>.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/vpath_pubs/8/
dc.identifier.articleid 1008
dc.identifier.contextkey 3783512
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath vpath_pubs/8
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/92508
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/vpath_pubs/8/2005_Gallup_NewQuickMethod.pdf|||Sat Jan 15 01:57:26 UTC 2022
dc.source.uri 10.1251/bpo107
dc.subject.disciplines Veterinary Pathology and Pathobiology
dc.subject.keywords Immunohistochemistry
dc.subject.keywords Sheep
dc.subject.keywords Gene expression
dc.subject.keywords Polymerase chain reaction
dc.title New quick method for isolating RNA from laser captured cells stained by immunofluorescent immunohistochemistry; RNA suitable for direct use in fluorogenic TaqMan one-step real-time RT-PCR
dc.type article
dc.type.genre article
dspace.entity.type Publication
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relation.isAuthorOfPublication 86c1ed73-b60d-48ce-8f35-449bc320a693
relation.isOrgUnitOfPublication cf38d7e3-b5f8-4859-83e3-ae8fab6a4c5f
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