-
Je něco špatně v tomto záznamu ?
Carboxymethylcellulose enhances the production of single-stranded DNA aptamers generated by asymmetric PCR
O. Redcenko, L. Draberova, P. Draber,
Jazyk angličtina Země Spojené státy americké
Typ dokumentu časopisecké články, práce podpořená grantem
Odkazy
PubMed
31704088
DOI
10.1016/j.ab.2019.113502
Knihovny.cz E-zdroje
- MeSH
- aptamerová technika SELEX metody MeSH
- aptamery nukleotidové chemie MeSH
- ELISA metody MeSH
- jednovláknová DNA chemie MeSH
- methylcelulosa chemie MeSH
- polymerázová řetězová reakce metody MeSH
- sodná sůl karboxymethylcelulosy chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nucleic acid aptamers are single-stranded (ss)DNA or RNA oligonucleotides that can take various conformations and bind specifically and with high affinity to selected targets. While the introduction of SELEX (systematic evolution of ligands by exponential enrichment) revolutionized the production of the aptamers, this procedure is impeded by the formation of undesirable by-products reflecting hybridization among complementary oligonucleotides in the ssDNA libraries during asymmetric PCR. To reduce nonspecific amplification we tested cellulose-derived compounds and found that sodium carboxymethylcellulose (CMC) at a concentration 0.05%-0.2% efficiently suppressed production of undesirable large DNA amplicons during asymmetric PCR in the course of SELEX. Formation of the PCR by-products was reduced by CMCs of low and medium viscosity more than by CMCs of high viscosity, and all of them bound to DNA oligonucleotides as determined by electrophoresis in agarose gels. In contrast to CMC, methylcellulose did not reduce the formation of the PCR by-products and did not bind to DNA. DNA aptamers selected in the presence of CMC could be used directly in enzyme-linked immunosorbent-like assay. The combined data suggest that CMC binds weekly to DNA oligonucleotides through hydroxyl groups and in this way inhibits low-affinity DNA-DNA hybridization and enhances the production of specific amplicons in asymmetric PCR.
- 000
- 00000naa a2200000 a 4500
- 001
- bmc20025267
- 003
- CZ-PrNML
- 005
- 20201222155135.0
- 007
- ta
- 008
- 201125s2020 xxu f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1016/j.ab.2019.113502 $2 doi
- 035 __
- $a (PubMed)31704088
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a xxu
- 100 1_
- $a Redcenko, Oleksij $u Department of Signal Transduction, Institute of Molecular Genetics of the Czech Academy of Sciences, CZ 14220, Prague, Czech Republic.
- 245 10
- $a Carboxymethylcellulose enhances the production of single-stranded DNA aptamers generated by asymmetric PCR / $c O. Redcenko, L. Draberova, P. Draber,
- 520 9_
- $a Nucleic acid aptamers are single-stranded (ss)DNA or RNA oligonucleotides that can take various conformations and bind specifically and with high affinity to selected targets. While the introduction of SELEX (systematic evolution of ligands by exponential enrichment) revolutionized the production of the aptamers, this procedure is impeded by the formation of undesirable by-products reflecting hybridization among complementary oligonucleotides in the ssDNA libraries during asymmetric PCR. To reduce nonspecific amplification we tested cellulose-derived compounds and found that sodium carboxymethylcellulose (CMC) at a concentration 0.05%-0.2% efficiently suppressed production of undesirable large DNA amplicons during asymmetric PCR in the course of SELEX. Formation of the PCR by-products was reduced by CMCs of low and medium viscosity more than by CMCs of high viscosity, and all of them bound to DNA oligonucleotides as determined by electrophoresis in agarose gels. In contrast to CMC, methylcellulose did not reduce the formation of the PCR by-products and did not bind to DNA. DNA aptamers selected in the presence of CMC could be used directly in enzyme-linked immunosorbent-like assay. The combined data suggest that CMC binds weekly to DNA oligonucleotides through hydroxyl groups and in this way inhibits low-affinity DNA-DNA hybridization and enhances the production of specific amplicons in asymmetric PCR.
- 650 _2
- $a aptamery nukleotidové $x chemie $7 D052157
- 650 _2
- $a sodná sůl karboxymethylcelulosy $x chemie $7 D002266
- 650 _2
- $a jednovláknová DNA $x chemie $7 D004277
- 650 _2
- $a ELISA $x metody $7 D004797
- 650 _2
- $a methylcelulosa $x chemie $7 D008747
- 650 _2
- $a polymerázová řetězová reakce $x metody $7 D016133
- 650 _2
- $a aptamerová technika SELEX $x metody $7 D052156
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Draberova, Lubica $u Department of Signal Transduction, Institute of Molecular Genetics of the Czech Academy of Sciences, CZ 14220, Prague, Czech Republic.
- 700 1_
- $a Draber, Petr $u Department of Signal Transduction, Institute of Molecular Genetics of the Czech Academy of Sciences, CZ 14220, Prague, Czech Republic. Electronic address: draberpe@img.cas.cz.
- 773 0_
- $w MED00000335 $t Analytical biochemistry $x 1096-0309 $g Roč. 589, č. - (2020), s. 113502
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/31704088 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y a $z 0
- 990 __
- $a 20201125 $b ABA008
- 991 __
- $a 20201222155131 $b ABA008
- 999 __
- $a ok $b bmc $g 1599412 $s 1115953
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2020 $b 589 $c - $d 113502 $e 20191105 $i 1096-0309 $m Analytical biochemistry $n Anal Biochem $x MED00000335
- LZP __
- $a Pubmed-20201125