Since only specific products contain this sequence, but none of the non-specific products, only specific products can be cloned. The other side of the linearized cloning vector has homology with a sequence present in the insert, but nested and non-overlapping with the gene-specific primer used for amplification. However, in contrast to ligation-independent cloning, the cloning vector has homology with only one of the two primers used for amplification of the insert. As with ligation-independent cloning, the strategy is based on homology between sequences present in both the vector and the insert. Biosci.We have developed an efficient strategy for cloning of PCR products that contain an unknown region flanked by a known sequence. Zhao G, Wei H and Guan Y 2013 Identification of a premature termination of DNA polymerization in vitro by Klenow fragment mutants. Zhou MY and Gomez-Sanchez CE 2000 Universal TA cloning. Zhao Y 2009 Construction of a high efficiency PCR Products cloning T vector using Avicenna. Zhao G and Guan Y 2010 Polymerization behavior of Klenow fragment and Taq DNA polymerase in short primer extension reactions. Zhang J, Li K, Pardinas JR, Sommer SS and Yao KT 2005 Proofreading genotyping assays mediated by high fidelity exo+ DNA polymerases. YunHua LU, LiXin MA and SiJing J 2006 A Universal high-throughput novel method of constructing the vectors. Yang YS, Watson WJ, Tucker PW and Capra JD 1993 Construction of recombinant DNA by exonuclease recession. Yang S, Li X, Ding D, Hou J, Jin Z, Yu X, Bo T, Li W and Li M 2005 A method for filling in the cohesive ends of double-stranded DNA using Pfu DNA polymerase. Stoker AW 1990 Cloning of PCR products after defined cohesive termini are created with T4 DNA polymerase. Ranjan RK and Rajagopal K 2010 Efficient ligation and cloning of DNA fragments with 2-bp overhangs. Motea EA and Berdis AJ 2010 Terminal deoxynucleotidyl transferase: The story of a misguided DNA polymerase. Liu X-P and Liu J-H 2010 The terminal 5 ' phosphate and proximate phosphorothioate promote ligation-independent cloning. Kaluz S, Kolble K and Reid KB 1992 Directional cloning of PCR products using exonuclease III. Hu G 1993 DNA polymerase-catalyzed addition of nontemplated extra nucleotides to the 3' end of a DNA fragment. Howland SW, Poh C-M and Renia L 2011 Directional, seamless, and restriction enzyme-free construction of random-primed complementary DNA libraries using phosphorothioate-modified primers. Biochemistry 30 1441–1448įreemont PS, Friedman JM, Beese LS, Sanderson MR and Steitz TA 1988 Cocrystal structure of an editing complex of Klenow fragment with DNA. 198 123–127Įger BT, Kuchta RD, Carroll SS, Benkovic PA, Dahlberg ME, Joyce CM and Benkovic SJ 1991 Mechanism of DNA replication fidelity for three mutants of DNA polymerase I: Klenow fragment KF(exo+), KF(polA5), and KF(exo-). 16 9677–9686Ĭlark JM, Joyce CM and Beardsley GP 1987 Novel blunt-end addition reactions catalyzed by DNA polymerase I of Escherichia coli. USA 91 10670–10674Ĭlark JM 1988 Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases. 247 241–248Ĭarver TE Jr, Hochstrasser RA and Millar DP 1994 Proofreading DNA: recognition of aberrant DNA termini by the Klenow fragment of DNA polymerase I. A proofreading function for the 3' leads to 5' exonuclease activity in deoxyribonucleic acid polymerases. 498 75–90īrutlag D and Kornberg A 1972 Enzymatic synthesis of deoxyribonucleic acid. EMBO J 10 25–33īerrow NS, Alderton D and Owens RJ 2009 The precise engineering of expression vectors using high-throughput In-Fusion PCR cloning. 402 203–205īeese LS and Steitz TA 1991 Structural basis for the 3'-5' exonuclease activity of Escherichia coli DNA polymerase I: a two metal ion mechanism. An Y, Wu W and Lv A 2010 A PCR-after-ligation method for cloning of multiple DNA inserts.
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