A facile method for the construction of synthetic genes |
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| Authors: | Pradhyuman K Singh Bijaya K Sarangi Rakesh Tuli |
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| Institution: | (1) Centre for Plant Molecular Biology, National Botanical Research Institute, Rana Pratap Marg, 226001 Lucknow, India |
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| Abstract: | A simple protocol for rapid assembly of chemically synthesized deoxyoligonucleotides into double stranded DNA is described.
Several parameters of a ligation-free method were investigated to allow efficient assembly of a large number of oligonucleotides
into double stranded DNA by polymerase chain reaction. Synthesis of a 701 bp DNA was carried out in a single reaction by assembling
28 oligonucleotides designed with partial overlaps at complementary ends. An estimate of error rate was made by sequencing
several independent clones of the synthesized DNA |
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| Keywords: | Gene synthesis oligonucleotide assembly polymerase chain reaction DNA assembly gene designing Pyrococcus polymerase Deep Vent polymerase |
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