Synthesis of long nucleic acid sequences
View Patent ↗The invention provides methods for the synthesis of long oligonucleotides, genes and gene fragments. The methods include the manufacture of genes or gene fragments that can be then inserted into a variety of vectors.
1. A method of synthesizing a gene block of a desired sequence greater than 100 bases, the method comprising:
a) providing a plurality of solid supports, wherein at least one solid support contains between 1-8% by weight controlled pore glass;
b) synthesizing oligonucleotides on the solid supports, wherein the oligonucleotides comprise a set of oligonucleotides that are partially complementary to at least one other oligonucleotide in the set, and the set of oligonucleotides as a group contain the desired sequence of the gene block;
c) detaching the oligonucleotides from the solid supports;
d) combining the oligonucleotides under conditions to synthesize the gene block.
2. The method of claim 1 wherein at least one solid support contains 5-8% by weight controlled pore glass.
3. The method of claim 1 wherein the gene block is combined with additional gene blocks to form a longer gene block.
4. The method of claim 1 further comprising performing an error correction step after combining the oligonucleotides.
5. The method of claim 1 wherein the gene block is greater than 200 bases.
6. The method of claim 1 wherein the gene block is greater than 400 bases.
7. The method of claim 1 wherein the gene block is greater than 500 bases.