Synthesis of long nucleic acid sequences
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.