Methods of cloning and producing fragment chains with readable information content

• US 7,371,851 B1
• Filed: 06/27/2000
• Issued: 05/13/2008
• Est. Priority Date: 03/18/1999
• Status: Expired due to Fees

• Current Assignee: Complete Genomics Incorporated (BGI Genomics Co., Ltd.)
• Original Assignee: Complete Genomics Incorporated (BGI Genomics Co., Ltd.)
• Inventors: Lexow, Preben
• Primary Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US10/019,258
• Time in Patent Office: 2,877 Days
• Field of Search: 435/6, 536/25.3, 536/23.1, 536/24.3
• US Class Current: 536/25.3
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups
• CPC Class Codes: C12N 15/66 General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The present invention provides a method of attaching a fragment of a first nucleic acid molecule to a second nucleic acid molecule using adapters to mediate the binding particularly in methods of cloning, methods of producing fragment chains with a readily readable information content, particularly comprising fragments corresponding to code, such as alphanumeric code, the nucleic acid molecules thus produced and kits for performing such methods.

Claims

1. A method for synthesizing a double stranded nucleic acid molecule that contains information that represents computer-readable binary code, comprising at least the steps of:

i) hybridizing together a plurality of double-stranded nucleic acid fragments, each fragment consisting of between 8 and 25 bases and comprising at least one sequence of between 4 and 10 bases that represent a unit of the binary code, and each fragment comprising at least one single stranded region that is capable of hybridizing to at least one other fragment; and

ii) optionally ligating the hybridized fragments;

to produce a double stranded nucleic acid molecule comprising a series of binary code units.

2. A method according to claim 1, wherein at least 10 double stranded nucleic acid fragments are hybridized together in step (i), to produce a double-stranded nucleic acid molecule comprising 10 fragments.

3. A method according to claim 1, wherein a plurality of double stranded nucleic acid molecules comprising a series of double-stranded nucleic acid fragments are synthesized and linked together.

4. A method of identifying at least one binary code unit contained within a double stranded nucleic acid molecule produced according to claim 1, comprising the steps of:

i) binding a labelled probe that is specific to at least one code unit to the unit; and

ii) detecting the label associated with the bound probe, thereby detecting the presence of the binary code unit to which the probe binds.