Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples

2.50
Hdl Handle:
http://hdl.handle.net/10147/239391
Title:
Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples
Authors:
Mee, Blanaid C.; Carroll, Paul; Donatello, Simona; Connolly, Elizabeth; Griffin, Mairead; Dunne, Barbara; Burke, Louise; Flavin, Richard; Rizkalla, Hala; Ryan, Ciara; Hayes, Brian; D'Adhemar, Charles; Banville, Niamh; Faheem, Nazia; Muldoon, Cian; Gaffney, Eoin F.
Citation:
Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples 2011, 9 (4):389 Biopreservation and Biobanking
Publisher:
Mary Ann Liebert Inc
Journal:
Biopreservation and Biobanking
Issue Date:
29-Dec-2011
URI:
http://hdl.handle.net/10147/239391
DOI:
10.1089/bio.2011.0034
Additional Links:
http://www.liebertonline.com/doi/abs/10.1089/bio.2011.0034
Abstract:
The Saint James's Hospital Biobank was established in 2008, to develop a high-quality breast tissue BioResource, as a part of the breast cancer clinical care pathway. The aims of this work were: (1) to ascertain the quality of RNA, DNA, and protein in biobanked carcinomas and normal breast tissues, (2) to assess the efficacy of AllPrep® (Qiagen) in isolating RNA, DNA, and protein simultaneously, (3) to compare AllPrep with RNEasy® and QIAamp® (both Qiagen), and (4) to examine the effectiveness of Allprotect® (Qiagen), a new tissue stabilization medium in preserving DNA, RNA, and proteins. One hundred eleven frozen samples of carcinoma and normal breast tissue were analyzed. Tumor and normal tissue morphology were confirmed by frozen sections. Tissue type, tissue treatment (Allprotect vs. no Allprotect), extraction kit, and nucleic acid quantification were analyzed by utilizing a 4 factorial design (SPSS PASW 18 Statistics Software®). QIAamp (DNA isolation), AllPrep (DNA, RNA, and Protein isolation), and RNeasy (RNA isolation) kits were assessed and compared. Mean DNA yield and A260/280 values using QIAamp were 33.2 ng/μL and 1.86, respectively, and using AllPrep were 23.2 ng/μL and 1.94. Mean RNA yield and RNA Integrity Number (RIN) values with RNeasy were 73.4 ng/μL and 8.16, respectively, and with AllPrep were 74.8 ng/μL and 7.92. Allprotect-treated tissues produced higher RIN values of borderline significance (P=0.055). No discernible loss of RNA stability was detected after 6 h incubation of stabilized or nonstabilized tissues at room temperature or 4°C or in 9 freeze-thaw cycles. Allprotect requires further detailed evaluation, but we consider AllPrep to be an excellent option for the simultaneous extraction of RNA, DNA, and protein from tumor and normal breast tissues. The essential presampling procedures that maintain the diagnostic integrity of pathology specimens do not appear to compromise the quality of molecular isolates.
Item Type:
Article
Language:
en
ISSN:
1947-5535; 1947-5543

Full metadata record

DC FieldValue Language
dc.contributor.authorMee, Blanaid C.en_GB
dc.contributor.authorCarroll, Paulen_GB
dc.contributor.authorDonatello, Simonaen_GB
dc.contributor.authorConnolly, Elizabethen_GB
dc.contributor.authorGriffin, Maireaden_GB
dc.contributor.authorDunne, Barbaraen_GB
dc.contributor.authorBurke, Louiseen_GB
dc.contributor.authorFlavin, Richarden_GB
dc.contributor.authorRizkalla, Halaen_GB
dc.contributor.authorRyan, Ciaraen_GB
dc.contributor.authorHayes, Brianen_GB
dc.contributor.authorD'Adhemar, Charlesen_GB
dc.contributor.authorBanville, Niamhen_GB
dc.contributor.authorFaheem, Naziaen_GB
dc.contributor.authorMuldoon, Cianen_GB
dc.contributor.authorGaffney, Eoin F.en_GB
dc.date.accessioned2012-08-22T08:46:20Z-
dc.date.available2012-08-22T08:46:20Z-
dc.date.issued2011-12-29-
dc.identifier.citationMaintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples 2011, 9 (4):389 Biopreservation and Biobankingen_GB
dc.identifier.issn1947-5535-
dc.identifier.issn1947-5543-
dc.identifier.doi10.1089/bio.2011.0034-
dc.identifier.urihttp://hdl.handle.net/10147/239391-
dc.description.abstractThe Saint James's Hospital Biobank was established in 2008, to develop a high-quality breast tissue BioResource, as a part of the breast cancer clinical care pathway. The aims of this work were: (1) to ascertain the quality of RNA, DNA, and protein in biobanked carcinomas and normal breast tissues, (2) to assess the efficacy of AllPrep® (Qiagen) in isolating RNA, DNA, and protein simultaneously, (3) to compare AllPrep with RNEasy® and QIAamp® (both Qiagen), and (4) to examine the effectiveness of Allprotect® (Qiagen), a new tissue stabilization medium in preserving DNA, RNA, and proteins. One hundred eleven frozen samples of carcinoma and normal breast tissue were analyzed. Tumor and normal tissue morphology were confirmed by frozen sections. Tissue type, tissue treatment (Allprotect vs. no Allprotect), extraction kit, and nucleic acid quantification were analyzed by utilizing a 4 factorial design (SPSS PASW 18 Statistics Software®). QIAamp (DNA isolation), AllPrep (DNA, RNA, and Protein isolation), and RNeasy (RNA isolation) kits were assessed and compared. Mean DNA yield and A260/280 values using QIAamp were 33.2 ng/μL and 1.86, respectively, and using AllPrep were 23.2 ng/μL and 1.94. Mean RNA yield and RNA Integrity Number (RIN) values with RNeasy were 73.4 ng/μL and 8.16, respectively, and with AllPrep were 74.8 ng/μL and 7.92. Allprotect-treated tissues produced higher RIN values of borderline significance (P=0.055). No discernible loss of RNA stability was detected after 6 h incubation of stabilized or nonstabilized tissues at room temperature or 4°C or in 9 freeze-thaw cycles. Allprotect requires further detailed evaluation, but we consider AllPrep to be an excellent option for the simultaneous extraction of RNA, DNA, and protein from tumor and normal breast tissues. The essential presampling procedures that maintain the diagnostic integrity of pathology specimens do not appear to compromise the quality of molecular isolates.en_GB
dc.language.isoenen
dc.publisherMary Ann Liebert Incen_GB
dc.relation.urlhttp://www.liebertonline.com/doi/abs/10.1089/bio.2011.0034en_GB
dc.rightsArchived with thanks to Biopreservation and Biobankingen_GB
dc.titleMaintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samplesen_GB
dc.typeArticleen
dc.identifier.journalBiopreservation and Biobankingen_GB
dc.description.provinceLeinsteren
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