Polymerase Chain Reaction and Transcription Using Locked Nucleic Acid Nucleotide Triphosphates

2008 ◽  
Vol 130 (26) ◽  
pp. 8124-8125 ◽  
Author(s):  
Rakesh N. Veedu ◽  
Birte Vester ◽  
Jesper Wengel
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Locked Nucleic Acid ◽  
Chain Reaction ◽  
Nucleotide Triphosphates ◽  
Polymerase Chain

scholarly journals Inhibition of microRNA-21 via locked nucleic acid-anti-miR suppressed metastatic features of colorectal cancer cells through modulation of programmed cell death 4

Tumor Biology ◽  
2017 ◽  
Vol 39 (3) ◽  
pp. 101042831769226 ◽  
Author(s):  
Reza Nedaeinia ◽  
Mohammadreza Sharifi ◽  
Amir Avan ◽  
Mohammad Kazemi ◽  
Abdolreza Nabinejad ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Polymerase Chain Reaction ◽  
Cell Death ◽  
Nucleic Acid ◽  
Locked Nucleic Acid ◽  
Chain Reaction ◽  
Programmed Cell Death 4 ◽  
Polymerase Chain ◽  
Novel Therapeutic

Colorectal cancer is among the most lethal of malignancies, due to its propensity to metastatic spread and multifactorial-chemoresistance. The latter property supports the need to identify novel therapeutic approaches for the treatment of colorectal cancer. MicroRNAs are endogenous non-coding small RNA molecules that function as post-transcriptional regulators of gene expression. Recently, programmed cell death 4 has been identified as a protein that increases during apoptosis. This gene is among the potential targets of miR-21 (OncomiR). Locked nucleic acid–modified oligonucleotides have recently emerged as a potential therapeutic option for targeting microRNAs. The aim of this study was to explore the functional role of locked nucleic acid-anti-miR-21 in the LS174T cell line in vitro and in vivo models. LS174T cells were treated with locked nucleic acid-anti-miR-21 for 24, 48, and 72 h in vitro. The expression of miR-21 and PDCD4 at messenger RNA (mRNA) level was evaluated by quantitative real-time polymerase chain reaction, while the protein level of PDCD4 was determined by Western blotting. Cell migratory behavior and the cluster-forming ability of cells were assessed before and after therapy. The disseminated tumor cells were assessed in the chick chorioallantoic membrane model by Alu quantitative polymerase chain reaction. Locked nucleic acid-anti-miR-21 was transfected successfully into the LS174T cells and inhibited the expression of miR-21. Locked nucleic acid-anti-miR-21 inhibited the migration and the number of cells forming clusters. Moreover, we found that locked nucleic acid-anti-miR-21 transfection was associated with a significant reduction in metastatic properties as assessed by the in ovo model. Our findings demonstrated the novel therapeutic potential of locked nucleic acid-anti-miR-21 in colon adenocarcinoma with high miR-21 expression.

scholarly journals Detection of the Janus kinase 2 V617F mutation using a locked nucleic-acid, real-time polymerase chain reaction assay

2018 ◽  
Vol 7 (1) ◽  
Author(s):  
Tshiphiri Senamela ◽  
Marleen Kock ◽  
Piet Becker ◽  
Joachim J.C. Potgieter
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Real Time ◽  
Jak2 V617f ◽  
Locked Nucleic Acid ◽  
Jak2 V617f Mutation ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain ◽  
V617f Mutation

The purpose of this study was to develop a real time polymerase chain reaction (PCR) assay for the detection of the JAK2 V617F mutation that could be used in diagnostic laboratories. Sanger sequencing and a newly developed locked nucleic-acid, real-time PCR assay were used to detect the JAK2 V617F mutation. There was 100% agreement between the sequencing and PCR analysis. Both assays were able to detect the mutation in all 24 of the 60 test specimens harbouring the mutation.

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