通过变性梯度凝胶电泳（DGGE）和化学变性梯度凝胶电泳（CDGE）联用提高富含鸟嘌呤-胞嘧啶（GC）的DNA片段中的突变检测
Improved mutation detection in GC-rich DNA fragments by combined DGGE and CDGE.
作者信息
Wu Y, Stulp R P, Elfferich P, Osinga J, Buys C H, Hofstra R M
机构信息
Department of Medical Genetics, University of Groningen, Antonius Deusinglaan 4, 9713 AW Groningen, The Netherlands.
出版信息
Nucleic Acids Res. 1999 Aug 1;27(15):e9. doi: 10.1093/nar/27.15.e9.
Abstract
Denaturing gradient gel electrophoresis (DGGE) has proven to be a powerful pre-screening method for the detection of DNA variants. If such variants occur, however, in DNA fragments that are very rich in G and C, they may escape detection. To overcome this limitation, we tested a novel gel system which combines DGGE and constant denaturant gel electrophoresis (CDGE), as it might have the advantages of both methods. Indeed, this combination had the advantages of both methods, good separation of hetero-duplex molecules and prevention of total strand dissociation, and it proved successful in the detection of DNA variants in several GC-rich fragments.
摘要
变性梯度凝胶电泳（DGGE）已被证明是一种用于检测DNA变异的强大预筛选方法。然而，如果此类变异出现在富含G和C的DNA片段中，它们可能会逃过检测。为了克服这一局限性，我们测试了一种新型凝胶系统，该系统结合了DGGE和恒定变性剂凝胶电泳（CDGE），因为它可能兼具两种方法的优点。事实上，这种组合兼具两种方法的优点，能很好地分离异源双链分子并防止链完全解离，并且在检测多个富含GC的片段中的DNA变异方面被证明是成功的。
相似文献
1
Improved mutation detection in GC-rich DNA fragments by combined DGGE and CDGE.通过变性梯度凝胶电泳（DGGE）和化学变性梯度凝胶电泳（CDGE）联用提高富含鸟嘌呤-胞嘧啶（GC）的DNA片段中的突变检测
Nucleic Acids Res. 1999 Aug 1;27(15):e9. doi: 10.1093/nar/27.15.e9.
2
RET and GDNF gene scanning in Hirschsprung patients using two dual denaturing gel systems.使用两种双重变性凝胶系统对先天性巨结肠患者进行RET和GDNF基因扫描。
Hum Mutat. 2000;15(5):418-29. doi: 10.1002/(SICI)1098-1004(200005)15:5<418::AID-HUMU3>3.0.CO;2-2.
3
Comparison of DGGE and CDGE in detection of single base changes in the hamster hprt and human N-ras genes.变性梯度凝胶电泳（DGGE）和恒变性凝胶电泳（CDGE）检测仓鼠次黄嘌呤磷酸核糖转移酶（hprt）基因和人类N- 原癌基因（N-ras）单碱基变化的比较
Mutat Res. 1995 Jun;334(3):357-64. doi: 10.1016/0165-1161(95)90073-x.
4
Detection of RET mutations in multiple endocrine neoplasia type 2a and familial medullary thyroid carcinoma by denaturing gradient gel electrophoresis.通过变性梯度凝胶电泳检测2a型多发性内分泌腺瘤病和家族性甲状腺髓样癌中的RET突变。
Hum Mutat. 1996;7(2):100-4. doi: 10.1002/(SICI)1098-1004(1996)7:2<100::AID-HUMU2>3.0.CO;2-G.
5
Mutation screening of the TP53 gene by temporal temperature gradient gel electrophoresis.通过时间温度梯度凝胶电泳对TP53基因进行突变筛查。
Methods Mol Biol. 2005;291:207-16. doi: 10.1385/1-59259-840-4:207.
6
Constant denaturant gel electrophoresis, a modification of denaturing gradient gel electrophoresis, in mutation detection.
Mutat Res. 1991 Jan;262(1):63-71. doi: 10.1016/0165-7992(91)90108-g.
7
Diagnosis of multiple endocrine neoplasia [MEN] 2A, 2B and familial medullary thyroid cancer [FMTC] by multiplex PCR and heteroduplex analyses of RET proto-oncogene mutations.通过多重聚合酶链反应和RET原癌基因突变的异源双链分析诊断多发性内分泌腺瘤病[MEN]2A、2B和家族性甲状腺髓样癌[FMTC]
Hum Mutat. 1996;8(1):64-70. doi: 10.1002/(SICI)1098-1004(1996)8:1<64::AID-HUMU9>3.0.CO;2-P.
8
The RET protooncogene in sporadic pheochromocytomas: frequent MEN 2-like mutations and new molecular defects.散发性嗜铬细胞瘤中的RET原癌基因：频繁出现的MEN 2样突变及新的分子缺陷
J Clin Endocrinol Metab. 1995 Jul;80(7):2063-8. doi: 10.1210/jcem.80.7.7608256.
9
Detection of mutations by denaturing gradient gel electrophoresis.通过变性梯度凝胶电泳检测突变
Curr Protoc Hum Genet. 2001 May;Chapter 7:Unit 7.5. doi: 10.1002/0471142905.hg0705s17.
10
DNA polymorphisms and conditions for SSCP analysis of the 20 exons of the ret proto-oncogene.
Oncogene. 1995 Mar 16;10(6):1257.
引用本文的文献
1
Frequency of germline PTEN mutations in differentiated thyroid cancer.分化型甲状腺癌中胚系 PTEN 突变的频率。
Thyroid. 2011 May;21(5):505-10. doi: 10.1089/thy.2010.0365. Epub 2011 Mar 21.
2
Balancing selection, sexual selection and geographic structure in MHC genes of Great Snipe.大沙锥MHC基因中的平衡选择、性选择和地理结构
Genetica. 2010 Apr;138(4):453-61. doi: 10.1007/s10709-008-9335-x. Epub 2008 Dec 4.
3
Large-scale mutation screening in patients with dilated or hypertrophic cardiomyopathy: a pilot study using DGGE.扩张型或肥厚型心肌病患者的大规模突变筛查：一项使用变性梯度凝胶电泳的初步研究
J Mol Med (Berl). 2006 Aug;84(8):682-91. doi: 10.1007/s00109-006-0056-2. Epub 2006 May 20.
4
Association of hereditary nonpolyposis colorectal cancer-related tumors displaying low microsatellite instability with MSH6 germline mutations.显示微卫星低度不稳定的遗传性非息肉病性结直肠癌相关肿瘤与MSH6种系突变的关联。
Am J Hum Genet. 1999 Nov;65(5):1291-8. doi: 10.1086/302612.
本文引用的文献
1
Improvement of fragment and primer selection for mutation detection by denaturing gradient gel electrophoresis.通过变性梯度凝胶电泳改进用于突变检测的片段和引物选择
Nucleic Acids Res. 1998 Dec 1;26(23):5432-40. doi: 10.1093/nar/26.23.5432.
2
Detection of mutations in GC-rich DNA by bisulphite denaturing gradient gel electrophoresis.通过亚硫酸氢盐变性梯度凝胶电泳检测富含鸟嘌呤-胞嘧啶的DNA中的突变。
Nucleic Acids Res. 1998 Mar 15;26(6):1548-9. doi: 10.1093/nar/26.6.1548.
3
Single-step DGGE-based mutation scanning of the p53 gene: application to genetic diagnosis of colorectal cancer.
Hum Mutat. 1997;9(4):348-55. doi: 10.1002/(SICI)1098-1004(1997)9:4<348::AID-HUMU8>3.0.CO;2-1.
4
MSH2 and MLH1 mutations in sporadic replication error-positive colorectal carcinoma as assessed by two-dimensional DNA electrophoresis.通过二维DNA电泳评估散发性复制错误阳性结直肠癌中的MSH2和MLH1突变。
Genes Chromosomes Cancer. 1997 Apr;18(4):269-78.
5
RET mutation screening in familial cutaneous lichen amyloidosis and in skin amyloidosis associated with multiple endocrine neoplasia.家族性皮肤苔藓样淀粉样变及与多发性内分泌肿瘤相关的皮肤淀粉样变中的RET突变筛查
J Invest Dermatol. 1996 Aug;107(2):215-8. doi: 10.1111/1523-1747.ep12329651.
6
Molecular analysis of phenylketonuria in Denmark: 99% of the mutations detected by denaturing gradient gel electrophoresis.丹麦苯丙酮尿症的分子分析：变性梯度凝胶电泳检测到的突变占99%。
Genomics. 1993 Jul;17(1):141-6. doi: 10.1006/geno.1993.1295.
7
Use of chemical clamps in denaturing gradient gel electrophoresis: application in the detection of the most frequent Mediterranean beta-thalassemic mutations.化学夹在变性梯度凝胶电泳中的应用：在检测最常见的地中海β-地中海贫血突变中的应用。
PCR Methods Appl. 1993 Oct;3(2):122-4. doi: 10.1101/gr.3.2.122.
8
Comparison of DGGE and CDGE in detection of single base changes in the hamster hprt and human N-ras genes.变性梯度凝胶电泳（DGGE）和恒变性凝胶电泳（CDGE）检测仓鼠次黄嘌呤磷酸核糖转移酶（hprt）基因和人类N- 原癌基因（N-ras）单碱基变化的比较
Mutat Res. 1995 Jun;334(3):357-64. doi: 10.1016/0165-1161(95)90073-x.
9
DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory.通过单碱基对替换而不同的DNA片段在变性梯度凝胶中被分离：与解链理论的对应关系。
Proc Natl Acad Sci U S A. 1983 Mar;80(6):1579-83. doi: 10.1073/pnas.80.6.1579.
10
Detection and localization of single base changes by denaturing gradient gel electrophoresis.通过变性梯度凝胶电泳检测和定位单碱基变化
Methods Enzymol. 1987;155:501-27. doi: 10.1016/0076-6879(87)55033-9.
Zotero 插件