منابع مشابه
Inverse Fusion PCR Cloning
Inverse fusion PCR cloning (IFPC) is an easy, PCR based three-step cloning method that allows the seamless and directional insertion of PCR products into virtually all plasmids, this with a free choice of the insertion site. The PCR-derived inserts contain a vector-complementary 5'-end that allows a fusion with the vector by an overlap extension PCR, and the resulting amplified insert-vector fu...
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We present a method that allows simultaneous fusion and cloning of multiple PCR products in a rapid and efficient manner. The procedure is based on the use of PCR primers that contain a single deoxyuridine residue near their 5' end. Treatment of the PCR products with a commercial deoxyuridine-excision reagent generates long 3' overhangs designed to specifically complement each other. The combin...
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We designed a new inverse PCR protocol combined with switching mechanism at 5' end of RNA transcript (SMART) technology, and applied it to the cloning of teleost corticotropin-releasing hormone precursor cDNA. Due to the advantages of both techniques, this method can efficiently amplify the complete 5'- and 3'-ends of cDNA in a single reaction, and might prove to be an alternative to the conven...
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B-cell functions as a key player in the humoral immunity producing immunoglobulin protein in mammalian. To produce wide-ranged and antigen-specific antibody, the B-cell undergoes genetic rearrangement of immunoglobulin genes during its maturation process. The genetic rearrangements in immunoglobulin genes require unstable steps for genome; breakage and re-ligation of the double strand DNA. In t...
متن کاملModified inverse PCR method for cloning the flanking sequences from human cell pools.
Inverse polymerase chain reaction (PCR) is a powerful approach for cloning the DNA sequences lying outside the boundaries of known sequences (7). This approach can be used to determine the integration site of a virus or a transposon (2,4). The procedure described by Triglia (7) can be directly used to amplify the flanking sequence in a plasmid or in a bacterial genome. However, it has been diff...
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ژورنال
عنوان ژورنال: PLoS ONE
سال: 2012
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0035407