directional cloning
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2020 ◽  
Vol 2020 (11) ◽  
pp. pdb.prot101238
Author(s):  
Michael R. Green ◽  
Joseph Sambrook

PLoS ONE ◽  
2017 ◽  
Vol 12 (11) ◽  
pp. e0186568
Author(s):  
Athanasios Niarchos ◽  
Anastasia Siora ◽  
Evangelia Konstantinou ◽  
Vasiliki Kalampoki ◽  
George Lagoumintzis ◽  
...  

2016 ◽  
Vol 13 (5) ◽  
pp. 1-2 ◽  
Author(s):  
Jens Allmer ◽  
Malik Yousef

EditorialThe term MicroRNA or its contraction miRNA currently appears in 21,215 titles of abstracts, published between 1997 and now, available on Pubmed (2016-21-22:12:59 EET). 4,108 of these were published in 2016 alone which signifies the importance of miRNA-related research. MicroRNAs can be detected experimentally using various techniques like directional cloning of endogenous small RNAs but they are time consuming [1]. Additionally, it is necessary for the miRNA and its mRNA target(s) to be co-expressed to infer a functional relationship which is difficult, if not impossible, to achieve [2]. Since experimental approaches are facing such difficulties, they have been complemented by computational approaches [3] thereby defining the field of computational miRNomics. Due to the rapid development in the discipline, it is important to assess the state-of-the-art. In this special issue, several areas of the field are investigated ranging from pre-miRNA detection via machine learning to application of differential expression analysis in plants. First, Saçar Demirci et al. discuss an approach to virus pre-miRNA detection using machine learning [4]. Such approaches are based on parameterization of miRNAs and Yousef et al. discuss how to select among such features [5]. A different computational perspective is provided by Kotipalli et al. who model the kinetics of miRNA genesis and targeting [6]. To fuel more refined future models for genesis and targeting, it is important to establish miRNA and target expression under varying conditions. Zhang et al. [7] and Kanke et al. [8] discuss two approaches to quantify miRNAs and other non-coding short RNAs. Diler et al., finally, discuss actual biological implications of differentially expressed miRNAs [9]. This special issue on computational miRNomics, thus, provides a trajectory from detection of pre-miRNAs to biological implications of differentially expressed miRNAs. Additional topics will be covered in the upcoming second volume of the special issue on computational miRNomics.


2014 ◽  
Vol 565 ◽  
pp. 3-8 ◽  
Author(s):  
Ji Gang Li ◽  
Guo Ying Han ◽  
Xiu Min Li ◽  
Jiao Jiao Sun ◽  
Ke Jing Song ◽  
...  

Directional cloning is a prerequisite for the construction of expression vectors in molecular biology laboratories. Although TA cloning is widely used to clone unmodified PCR (polymerase chain reaction) products, a major disadvantage of this technique is that cloning is not directional. Here we reported a novel PCR products cloning vector with one deoxythymidine overhang and one deoxycytidine overhang at two 3'-ends respectively. With the choice of nucleotides of 5'-ends of PCR primers, PCR products can be cloned to this vector both directly and directionally. The feasibility and efficacy of this cloning method were confirmed by using a pET-17b derivative vector and a green fluorescent protein gene (EGFP) and a red fluorescent protein reporter (Ds-Red) gene. This cloning strategy may be useful in the high-throughput construction of expression vectors and could be viewed as an interesting improvement of existing TA cloning method.


2013 ◽  
Vol 38 (5) ◽  
pp. 857-866 ◽  
Author(s):  
Guojie Zhao ◽  
Jun Li ◽  
Tianyu Hu ◽  
Hua Wei ◽  
Yifu Guan

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