| 主权项 |
1. A method for producing a DNA sequence for a nucleic acid library comprising a plurality of elements,
wherein the DNA sequence comprises at least three sections, wherein the first and the third sections in each element have defined sequences that are identical in each element of the nucleic acid library, wherein a central second section comprises at least two adjoining variable codon triplets, wherein at least two oligonucleotide libraries and at least two single stranded oligonucleotides with the following characteristics are used in this method:
a first oligonucleotide library consisting of oligonucleotides each comprising a partial sequence corresponding to the defined first section of the nucleic acid library, at least one first variable codon triplet, a subsequently arranged first partial sequence of a spacer element and a defined 3′ overhang;a first complementary oligonucleotide or a first complementary oligonucleotide library, wherein the first complementary oligonucleotide is substantially complementary to the oligonucleotides of the first oligonucleotide library, comprising a partial sequence that is complementary to the first partial sequence of the spacer element of the oligonucleotides of the first oligonucleotide library, at least one first complementary variable or substantially complementary variable codon triplet and a partial sequence complementary to the first section of the oligonucleotides of the first oligonucleotide library;a second oligonucleotide library consisting of oligonucleotides each comprising a partial sequence forming the second partial sequence of the spacer element, at least one second variable codon triplet and a partial sequence corresponding to the defined second section of the nucleic acid library; anda second complementary oligonucleotide or a second complementary oligonucleotide library, wherein the second complementary oligonucleotide is substantially complementary to the oligonucleotides of the second oligonucleotide library comprising a partial sequence complementary to the second section of the second oligonucleotide library, at least one second complementary variable or substantially complementary variable codon triplet, a partial sequence complementary to the second partial sequence of the spacer element of the oligonucleotides of the second oligonucleotide library and a subsequently arranged defined 3′ overhang; OR
a first oligonucleotide library consisting of oligonucleotides each comprising a partial sequence corresponding to the defined first section of the nucleic acid library, at least one first variable codon triplet and a subsequently arranged first partial sequence of a spacer element;a first complementary oligonucleotide or a first substantially complementary oligonucleotide library, wherein the first complementary oligonucleotide is substantially complementary to the oligonucleotides of the first oligonucleotide library comprising a defined 5′ overhang, a partial sequence complementary to the first partial sequence of the spacer element of the oligonucleotides of the first oligonucleotide library, at least one first complementary variable or substantially complementary variable codon triplet and a partial sequence complementary to the first section of the oligonucleotides of the first oligonucleotide library;a second oligonucleotide library consisting of oligonucleotides with a defined 5′ overhang comprising a partial sequence forming the second partial sequence of the spacer element, at least one second variable codon triplet and a partial sequence corresponding to the defined second section of the nucleic acid library; anda second complementary oligonucleotide or a second substantially complementary oligonucleotide library, wherein the second complementary oligonucleotide is substantially complementary to the oligonucleotides of the second oligonucleotide library comprising a partial sequence complementary to the second section of the second oligonucleotide library, at least one second complementary variable or substantially complementary variable codon triplet, and a partial sequence complementary to the second partial sequence of the spacer element of the oligonucleotides of the second oligonucleotide library;the method comprising:
hybridizing the oligonucleotides of the first oligonucleotide library to the first substantially complementary oligonucleotide to generate a first double-stranded DNA library and hybridization of the oligonucleotides of the second oligonucleotide library to the second substantially complementary oligonucleotide to generate a second double-stranded DNA library; ligating the first double-stranded DNA library to the second double-stranded DNA library, thereby producing a library of double-stranded DNA ligation products, the library of double-stranded DNA ligation products comprising the following subsections:
a defined first section of the nucleic acid library,at least one first variable codon triplet,a spacer element,at least one second variable codon triplet; anda defined second section of the nucleic acid library; and removing the spacer element from the library of double-stranded DNA ligation products, thereby producing a library of double-stranded DNA-product comprising the following subsections:
a defined first section of the nucleic acid library,at least one first variable codon triplet,at least one second variable codon triplet; anda defined second section of the nucleic acid library. |
