dc.contributor.author |
Tolnai Zoltán |
|
dc.contributor.author |
Harkai Ákos |
|
dc.contributor.author |
Szeitner Zsuzsanna |
|
dc.contributor.author |
Nagyné Scholz Éva |
|
dc.contributor.author |
Percze Krisztina |
|
dc.contributor.author |
Gyurkovics Anna |
|
dc.contributor.author |
Mészáros Tamás |
|
dc.date.accessioned |
2022-06-14T09:44:07Z |
|
dc.date.available |
2022-06-14T09:44:07Z |
|
dc.date.issued |
2019 |
|
dc.identifier |
85054725862 |
|
dc.identifier.citation |
journalVolume=1047;journalTitle=ANALYTICA CHIMICA ACTA;pagerange=225-230;journalAbbreviatedTitle=ANAL CHIM ACTA; |
|
dc.identifier.uri |
http://repo.lib.semmelweis.hu//handle/123456789/6821 |
|
dc.identifier.uri |
doi:10.1016/j.aca.2018.10.017 |
|
dc.description.abstract |
Although various methods have been developed to suffice the oligonucleotide demand of molecular biology laboratories, in vitro production of high-purity ssDNAs remains to be a challenging task. We hypothesized that complementing the asymmetric PCR with 3’ phosphate blocked limiting primer decreases the mispriming thus reduces polymerisation of DNA by-products. The presented results attest our assumption that the primer blocked asymmetric PCR (PBA-PCR) selectively produces ssDNA of interest and is even suitable for effective amplification of DNA libraries of large sequence space. The high-throughput sequence analysis demonstrated that PBA-PCR also alleviates the PCR bias obstacle since it does not distort the sequence space. The practicability of the novel method was verified by monitoring the process of SELEX and screening of aptamer candidates using PBA-PCR produced ssDNAs in Amplified Luminescent Proximity Homogeneous Assay. In summary, we have developed a generally applicable method for straightforward, cost-effective production of ssDNA with on demand labelling. © 2018 Elsevier B.V. |
|
dc.format.extent |
225-230 |
|
dc.relation.ispartof |
urn:issn:0003-2670 |
|
dc.title |
A simple modification increases specificity and efficiency of asymmetric PCR |
|
dc.type |
Journal Article |
|
dc.date.updated |
2019-03-01T15:06:22Z |
|
dc.language.rfc3066 |
en |
|
dc.rights.holder |
NULL |
|
dc.identifier.mtmt |
30309501 |
|
dc.identifier.wos |
000453384400025 |
|
dc.identifier.pubmed |
30567654 |
|
dc.contributor.institution |
Növénybiológiai Intézet |
|
dc.contributor.institution |
Orvosi Vegytani, Molekuláris Biológiai és Patobiokémiai Intézet |
|
dc.contributor.institution |
MTA-BME Műszaki Analitikai Kémiai Kutatócsoport |
|
dc.contributor.institution |
MTA-SE Pathobiokémiai Kutatócsoport |
|
dc.contributor.institution |
Szervetlen és Analitikai Kémia Tanszék |
|