PMID- 20238183
OWN - NLM
STAT- MEDLINE
DCOM- 20100810
LR  - 20181113
IS  - 1559-0305 (Electronic)
IS  - 1073-6085 (Linking)
VI  - 45
IP  - 2
DP  - 2010 Jun
TI  - Temperature-assisted cyclic hybridization (TACH): an improved method for 
      supercoiled DNA hybridization.
PG  - 171-9
LID - 10.1007/s12033-010-9261-x [doi]
AB  - Accurate hybridization is dependent on the ratio between sequence-specific and 
      unspecific binding. Dissociation of unspecifically bound, while maintaining 
      specifically hybridized, nucleic acids are key steps to obtain a well-defined 
      complex. We have developed a new method, temperature-assisted, cyclic 
      hybridization (TACH), which increases cognate binding at the expense of 
      unspecific hybridization. The method was used for optimizing binding of peptide 
      nucleic acid (PNA) to supercoiled plasmids and has several advantages over 
      previous methods: (1) it reduces the required amount of bis-PNA by three- to 
      fourfold; (2) it results in less unspecific binding; (3) it extends cooperative 
      hybridization, from 3 bp to 5 bp between two adjacent binding sites; and (4) it 
      decreases the aggregation of bis-PNA. This method might be extended to other 
      forms of hybridizations including the use of additional nucleic acids analogs, 
      such as locked nucleic acid (LNA) and, also, to other areas where PNAs are used 
      such as fluorescence in situ hybridization (FISH), microarrays, or in vivo 
      plasmid delivery.
FAU - Oprea, Iulian I
AU  - Oprea II
AD  - Department of Laboratory Medicine, Clinical Research Center, Karolinska 
      Institutet, Karolinska University Hospital Huddinge, SE-141 86 Huddinge, Sweden. 
      iulian.oprea@ki.se
FAU - Simonson, Oscar E
AU  - Simonson OE
FAU - Moreno, Pedro M D
AU  - Moreno PM
FAU - Viola, Joana R
AU  - Viola JR
FAU - Lundin, Karin E
AU  - Lundin KE
FAU - Smith, C I Edvard
AU  - Smith CI
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Switzerland
TA  - Mol Biotechnol
JT  - Molecular biotechnology
JID - 9423533
RN  - 0 (DNA, Superhelical)
RN  - 0 (Peptide Nucleic Acids)
SB  - IM
MH  - Analysis of Variance
MH  - Animals
MH  - DNA, Superhelical/*chemistry/metabolism
MH  - Electrophoresis, Agar Gel
MH  - Gene Expression
MH  - Hydrogen-Ion Concentration
MH  - Mice
MH  - Nucleic Acid Conformation
MH  - Nucleic Acid Hybridization/*methods
MH  - Peptide Nucleic Acids/chemistry/metabolism
MH  - Plasmids/*chemistry/metabolism
MH  - Sensitivity and Specificity
MH  - Temperature
EDAT- 2010/03/20 06:00
MHDA- 2010/08/11 06:00
CRDT- 2010/03/19 06:00
PHST- 2010/03/19 06:00 [entrez]
PHST- 2010/03/20 06:00 [pubmed]
PHST- 2010/08/11 06:00 [medline]
AID - 10.1007/s12033-010-9261-x [doi]
PST - ppublish
SO  - Mol Biotechnol. 2010 Jun;45(2):171-9. doi: 10.1007/s12033-010-9261-x.