Optimized conditions for cloning PCR products into an XcmI T-vector.
نویسندگان
چکیده
products. BioTechniques 12:28-30. 10.Scharf, S.J., G.T. Horn and H.A. Erlich. 1986. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science 233:1076-1078. 11.Shuldiner, A.R., L.A. Scott and J. Roth. 1990. PCR-induced (ligase free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) product. Nucleic Acids Res. 18:1920. 12.Starr, L. and V. Quaranta. 1992. An efficient and reliable method for cloning PCRamplification products: a survey of point mutations in integrin cDNA. BioTechniques 13:612-618.
منابع مشابه
XcmI-containing vector for direct cloning of PCR products.
Many different strategies are used for cloning polymerase chain reaction (PCR) products. Some use restriction sites pre-integrated into primers or contained in a generated fragment. Others, such as the one used by Stratagene (La Jolla, CA, USA) in its pCR-Script Direct SK(+) Cloning Kit, are based on a blunt-end ligation. These strategies require the use of additional enzymes to polish the end...
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1.Atcheson, C.L., B. DiDomenico, S. Frackman, R.E. Esposito and R.T. Elder. 1987. Isolation, DNA sequence, and regulation of a meiosis-specific eukaryotic recombination gene. Proc. Natl. Acad. Sci. USA 84:80358039. 2.Cha, J., W. Bishai and S. Chandrasegaran. 1993. New vectors for direct cloning of PCR products [published erratum appears in Gene 1994; 141:149]. Gene 136:369-370. 3.Clark, J.M. 19...
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An improved procedure for preparing PCR cloning vectors was developed. This procedure includes the incorporation of adapters to create XcmI restriction enzyme sites in pBluescript II SK(+) vectors, digestion with XcmI followed by further digestion of the small fragment produced by XcmI digestion with additional enzymes, and purification with PCR purification kits. Using this procedure, PCR clon...
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The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3' adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3' overhanging thymidine. The aim of this work was to offer in a single vector the ability to clone both types of PCR products. For that p...
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ورودعنوان ژورنال:
- BioTechniques
دوره 22 1 شماره
صفحات -
تاریخ انتشار 1997