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首页> 外文期刊>Biopreservation and biobanking >Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples
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Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples

机译：保持乳腺癌样本的完整性以及从Allprotect稳定和不稳定的组织样品中单独或同时提取高质量的DNA，RNA和蛋白质

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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 A_(260/280) values using QIAamp were 33.2ng/μL and 1.86, respectively, and using AllPrep were 23.2ng/μL and 1.94. Mean RNA yield and RNA Integrity Number (RIN) values with RNeasy were 73.4ng/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℃ 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 pre- sampling procedures that maintain the diagnostic integrity of pathology specimens do not appear to compromise the quality of molecular isolates.
机译：圣詹姆斯医院生物库成立于2008年，目的是开发高质量的乳腺组织BioResource，作为乳腺癌临床护理途径的一部分。这项工作的目的是：（1）确定生物库癌和正常乳腺组织中RNA，DNA和蛋白质的质量，（2）评估AllPrep的功效。 （Qiagen）同时分离RNA，DNA和蛋白质，（3）比较AllPrep和RNEasy？和QIAamp？ （均为Qiagen），以及（4）检查Allprotect的有效性？ （Qiagen），一种用于保存DNA，RNA和蛋白质的新型组织稳定培养基。分析了一百一十一份癌和正常乳腺组织的冷冻样品。冷冻切片证实肿瘤和正常组织形态。通过使用四因子设计（SPSS PASW 18 Statistics Software？）分析组织类型，组织处理（Allprotect对比NoAllprotect），提取试剂盒和核酸定量。评估并比较了QIAamp（DNA分离），AllPrep（DNA，RNA和蛋白质分离）和RNeasy（RNA分离）试剂盒。使用QIAamp的平均DNA产量和A_（260/280）值分别为33.2ng /μL和1.86，使用AllPrep的平均DNA产量为23.2ng /μL和1.94。 RNeasy的平均RNA产量和RNA完整性数（RIN）值分别为RNeasy的73.4ng /μL和8.16，而AllPrep的平均值为74.8 ng /μL和7.92。经所有保护剂处理的组织产生的RIN值具有较高的临界意义（P = 0.055）。在室温或4℃或9次冻融循环中，将稳定或不稳定的组织孵育6小时后，没有发现可辨别的RNA稳定性损失。 Allprotect需要进一步详细评估，但是我们认为AllPrep是从肿瘤和正常乳腺组织中同时提取RNA，DNA和蛋白质的绝佳选择。维持病理标本诊断完整性的基本预采样程序似乎并未损害分子分离物的质量。

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