A general one-step method for the cloning of PCR products. Export

@article{citeulike:1828220, abstract = {A very fast, highly efficient, versatile and low-cost cloning of PCR products is described. PCR amplicons, obtained with any set of primers, is directly integrated into circular plasmid vectors by means of a one-step restriction-ligation procedure. When using proof-reading DNA polymerases, 100\% cloning efficiency is easily achieved, implying that direct cloning into 'final-use' vectors (i.e. avoiding any intermediate cloning step) is a feasible task. Albeit with a lower efficiency, the same procedure is also suitable for the cloning of PCR products generated by 'non-proof-reading' DNA polymerases. Furthermore, with a simple modification of the vector polylinker site, the present method can be easily adapted to the directional cloning of open-reading-frame-encoding amplicons. This one-step procedure thus couples high efficiency with high reliability and versatility, and lends itself as the method of choice for routine cloning of PCR products.}, address = {Dipartimento di Biochimica e Biologia Molecolare, Universit\`{a} degli Studi di Parma, Parco Area delle Scienze 23/A, Parma I-43100, Italy. angelo.bolchi@unipr.it}, author = {Bolchi, A. and Ottonello, S. and Petrucco, S.}, citeulike-article-id = {1828220}, citeulike-linkout-0 = {http://dx.doi.org/10.1042/BA20050050}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/15992357}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=15992357}, doi = {10.1042/BA20050050}, issn = {0885-4513}, journal = {Biotechnol Appl Biochem}, keywords = {blunt\_ligation, cloning}, month = {December}, number = {Pt 3}, pages = {205--209}, posted-at = {2009-04-25 02:27:00}, priority = {0}, title = {A general one-step method for the cloning of PCR products.}, url = {http://dx.doi.org/10.1042/BA20050050}, volume = {42}, year = {2005} }

TY - JOUR ID - citeulike:1828220 L3 - citeulike-article-id:1828220 N2 - A very fast, highly efficient, versatile and low-cost cloning of PCR products is described. PCR amplicons, obtained with any set of primers, is directly integrated into circular plasmid vectors by means of a one-step restriction-ligation procedure. When using proof-reading DNA polymerases, 100% cloning efficiency is easily achieved, implying that direct cloning into 'final-use' vectors (i.e. avoiding any intermediate cloning step) is a feasible task. Albeit with a lower efficiency, the same procedure is also suitable for the cloning of PCR products generated by 'non-proof-reading' DNA polymerases. Furthermore, with a simple modification of the vector polylinker site, the present method can be easily adapted to the directional cloning of open-reading-frame-encoding amplicons. This one-step procedure thus couples high efficiency with high reliability and versatility, and lends itself as the method of choice for routine cloning of PCR products. IS - Pt 3 JF - Biotechnol Appl Biochem SN - 0885-4513 AD - Dipartimento di Biochimica e Biologia Molecolare, Università degli Studi di Parma, Parco Area delle Scienze 23/A, Parma I-43100, Italy. angelo.bolchi@unipr.it EP - 209 TI - A general one-step method for the cloning of PCR products. VL - 42 SP - 205 KW - blunt_ligation KW - cloning AU - Bolchi, A AU - Ottonello, S AU - Petrucco, S PY - 2005/12// UR - http://dx.doi.org/10.1042/BA20050050 ER -