发明名称 |
COMPOSITIONS AND METHODS FOR HIGH FIDELITY ASSEMBLY OF NUCLEIC ACIDS |
摘要 |
<p>Aspects of the invention relate to methods, compositions and algorithms for designing and producing a target nucleic acid. The method can include: (1) providing a plurality of blunt-end double-stranded nucleic acid fragments having a restriction enzyme recognition sequence at both ends thereof; (2) producing via enzymatic digestion a plurality of cohesive-end double-stranded nucleic acid fragments each having two different and non-complementary overhangs; (3) ligating the plurality of cohesive-end double-stranded nucleic acid fragments with a ligase; and (4) forming a linear arrangement of the plurality of cohesive-end double-stranded nucleic acid fragments, wherein the unique arrangement comprises the target nucleic acid. In certain embodiments, the plurality of blunt-end double-stranded nucleic acid fragments can be provided by: releasing a plurality of oligonucleotides synthesized on a solid support; and synthesizing complementary strands of the plurality of oligonucleotides using a polymerase based reaction.</p> |
申请公布号 |
EP2748318(A2) |
申请公布日期 |
2014.07.02 |
申请号 |
EP20120753894 |
申请日期 |
2012.08.23 |
申请人 |
GEN9 INC. |
发明人 |
JACOBSON, JOSEPH;SCHINDLER, DANIEL;LAWTON, SCOTT S. |
分类号 |
C12N15/10;C12N15/66;C12P19/34;C40B50/06;G06F19/22 |
主分类号 |
C12N15/10 |
代理机构 |
|
代理人 |
|
主权项 |
|
地址 |
|