Amplification of GC-rich ADAMTS-2 and URG4/URGCP promoter regions with optimized combination of PCR enhancers

dc.contributor.authorAlper, Meltem
dc.contributor.authorTokay, Esra
dc.contributor.authorKöçkar, Feray
dc.date.accessioned2019-06-18T12:27:33Z
dc.date.available2019-06-18T12:27:33Z
dc.date.issued2016
dc.departmentTeknik Bilimler Meslek Yüksekokulu
dc.descriptionAlper, Meltem (Aksaray, Yazar)
dc.description.abstractPCR is an effective and widely used method for the amplification of target DNA fragments in vitro. The templates that have high guanine-cytosine (GC) content, complex secondary structures, or short tandem repeats are difficult to amplify by the conventional PCR methods. Human genome regulatory regions such as promoters are rich in terms of GC base compositions. To amplify these regions some special modifications or additives are required. Using different PCR strategies such as hot-start/touchdown PCR or the utilization of various additives into the reaction mixture such as organic molecules can improve PCR yield. In the present study we optimized the PCR conditions for the amplification of the human ADAMTS-2 gene promoter region featuring an extremely high GC content and a secondary structure. We show that a combination of three additives, betaine, dimethyl sulfoxide, and 7-deaza GTP, is essential to obtain a correct PCR amplicon. To demonstrate whether the optimized PCR condition was applicable to other GC-enriched regions, a similar procedure was repeated for the amplification of the human URG4/URGCP gene promoter.
dc.identifier.doi10.3906/biy-1502-37
dc.identifier.endpage205en_US
dc.identifier.issue1en_US
dc.identifier.scopusqualityQ1
dc.identifier.startpage196en_US
dc.identifier.urihttps:/dx.doi.org/10.3906/biy-1502-37
dc.identifier.urihttps://hdl.handle.net/20.500.12451/1403
dc.identifier.volume40en_US
dc.identifier.wosqualityN/A
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakTR-Dizin
dc.language.isoen
dc.publisherTurkish Journal of Biology
dc.relation.ispartofTurkish Journal of Biology
dc.relation.publicationcategoryMakale - Ulusal Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectGC-Rich PCR
dc.subjectPCR Enhancers
dc.subjectPromoter Amplifications
dc.subjectRobust PCR
dc.subjectGC Bakımından Zengin PCR
dc.subjectPCR Geliştiricileri
dc.subjectPromoter Yükseltmeleri
dc.subjectGüçlü PCR
dc.titleAmplification of GC-rich ADAMTS-2 and URG4/URGCP promoter regions with optimized combination of PCR enhancers
dc.typeArticle

Dosyalar

Orijinal paket
Listeleniyor 1 - 1 / 1
Yükleniyor...
Küçük Resim
İsim:
alper-meltem-2016.pdf
Boyut:
2.55 MB
Biçim:
Adobe Portable Document Format
Açıklama:
Tam Metin / Full Text
Lisans paketi
Listeleniyor 1 - 1 / 1
[ X ]
İsim:
license.txt
Boyut:
1.44 KB
Biçim:
Item-specific license agreed upon to submission
Açıklama: