TY - JOUR
T1 - An efficient method for generation and subcloning of tandemly repeated DNA sequences with defined length, orientation and spacing
AU - Jiang, Shi Wen
AU - Trujillo, Miguel A.
AU - Eberhardt, Norman L.
PY - 1996
Y1 - 1996
N2 - Tandemly repeated DNA sequences generated from single synthetic oligonucleotide monomers are useful for many purposes, With conventional ligation procedures low yields and random orientation of oligomers makes cloning of defined repeated sequences difficult, We solved these problems using 2 bp overhangs to direct orientation and random incorporation of linkers containing restriction sites during ligation. Ligation products are amplified by PCR using the linker oligonucleotides as primers. Restriction digestion of the PCR products generate multimer distributions whose length is controlled by the monomer/linker ratio, The concatenated DNA fragments of defined length, orientation and spacing can be directly used for subcloning or other applications without further treatment.
AB - Tandemly repeated DNA sequences generated from single synthetic oligonucleotide monomers are useful for many purposes, With conventional ligation procedures low yields and random orientation of oligomers makes cloning of defined repeated sequences difficult, We solved these problems using 2 bp overhangs to direct orientation and random incorporation of linkers containing restriction sites during ligation. Ligation products are amplified by PCR using the linker oligonucleotides as primers. Restriction digestion of the PCR products generate multimer distributions whose length is controlled by the monomer/linker ratio, The concatenated DNA fragments of defined length, orientation and spacing can be directly used for subcloning or other applications without further treatment.
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U2 - 10.1093/nar/24.16.3278
DO - 10.1093/nar/24.16.3278
M3 - Article
C2 - 8774914
AN - SCOPUS:0029781175
SN - 0305-1048
VL - 24
SP - 3278
EP - 3279
JO - Nucleic acids research
JF - Nucleic acids research
IS - 16
ER -