A universal method for directional cloning of PCR products based on asymmetric PCR

2009 ◽  
Vol 52 (1) ◽  
pp. 41 ◽  
Author(s):  
Bao-Li Wang ◽  
Yan-Li Jiao ◽  
Xiao-Xia Li ◽  
Fang Zheng ◽  
Hui Liang ◽  
...  
1999 ◽  
Vol 260 (6) ◽  
pp. 569-573 ◽  
Author(s):  
J. Gál ◽  
R. Schnell ◽  
S. Szekeres ◽  
M. Kálmán

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4140
Author(s):  
Gina L. Costa ◽  
Michael P. Weiner

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.


2009 ◽  
pp. NA-NA ◽  
Author(s):  
Maren Mix ◽  
Thomas Reske ◽  
Heiko Duwensee ◽  
Gerd-Uwe Flechsig

1996 ◽  
Vol 29 (2) ◽  
pp. 174-181 ◽  
Author(s):  
M. Boysen ◽  
Marisé Borja ◽  
Catalina del Moral ◽  
Oscar Salazar ◽  
V. Rubio

1992 ◽  
Vol 20 (16) ◽  
pp. 4369-4370 ◽  
Author(s):  
Stefan Kaluz ◽  
Konrad Kölble ◽  
Kenneth B.M. Reid

1996 ◽  
Vol 29 (2) ◽  
pp. 174-181 ◽  
Author(s):  
Marianne Boysen ◽  
Marisé Borja ◽  
Catalina del Moral ◽  
Oscar Salazar ◽  
Victor Rubio

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