Direct Ligation of Human CD4 Polymerase Chain Reaction Fragment into Vectors at Specific Restriction Sites with Positional Heterostagger Cloning
✍ Scribed by Ferenc Felfoldi; Zoltan Kupihar; Sandor Bottka; Laszlo G. Puskas
- Publisher
- Elsevier Science
- Year
- 1997
- Tongue
- English
- Weight
- 73 KB
- Volume
- 253
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
275
cently, BAC clones have gained recognition as the most or with the help of restriction enzymes, whose recognition sites were previously incorporated into the 5-end promising source of DNA for the commencement of largescale human genome sequencing. The strategy that has of the primers (2). A variety of other methods have been developed for making the cloning procedure more been envisaged involves, as a first step, the generation of end sequences from BACs representing 201 coverage efficient and faster such as the use of uracil DNA glycosylase (3), ligation-independent cloning (4), TA cloning of the human genome (11). The single-primer PCR approach, which can eliminate the purification and manip-(5, 6), and heterostagger cloning (7). Although these methods obviously possess several advantages, they ulation step, could be one of the most appropriate strategies for this kind of large-scale projects.
need special vectors and in some cases treatment of PCR products is necessary. The greatest drawback of Acknowledgments. This work was supported by the U.S. Departthese methods is, however, that they are not suitable ment of Energy (under Contract DE-AC05-96OR22464 with Lockheed-Martin Energy Systems, Inc.).