Compositions and methods for synthesizing nucleic acids

• US 20040126789A1
• Filed: 09/05/2003
• Published: 07/01/2004
• Est. Priority Date: 09/05/2002
• Status: Abandoned Application

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Invitrogen Corp. (Thermo Fisher Scientific Incorporated)
• Inventors: Park, Kyusung, Lee, Jun E.
• Application Number:
  US10/655,579
• Publication Number:
  US 20040126789A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: C12Q 2522/101 Single or double stranded nucleic acid binding proteins
• CPC Class Codes: C12Q 2527/127 the enzyme inhibitor or activator used
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin

Abstract

Disclosed are compositions preferably for use in nucleic acid synthesis that include one or more anti-reverse transcriptase (RT) antibodies and/or one or more anti-DNA polymerase (DNAP) antibodies and/or single strand binding proteins (SSBs). Some of the disclosed compositions include one or more anti-DNAP antibodies and/or one or more anti-RT antibodies and one or more SSBs. Other disclosed compositions include two or more SSBs. The disclosed nucleic acid synthesis compositions also can include one or more DNAPs, one or more RTs, one or more nucleotides, one or more primers, and/or one or more templates.

Also disclosed are methods for using such compositions in nucleic acid synthesis, amplification and sequencing, where various combinations of anti-RT antibodies, anti-DNAP antibodies and/or SSBs can improve the yield and/or homogeneity of primer extension products.

Claims

1. A composition comprising:

(a) at least one anti-DNAP antibody and/or at least one anti-RT antibody; and

(b) at least one SSB.

3. The composition of claim 1 or claim 2, further comprising one or more nucleic acid templates.

4. The composition of claim 3, wherein at least one of said one or more templates is a cDNA.

5. The composition of claim 1 or claim 2, further comprising at least one primer.

6. The composition of claim 1 or claim 2, further comprising one or more nucleotides.

7. The composition of claim 1 or claim 2, further comprising one or more DNAPs.

8. The composition of claim 1 or claim 2, further comprising one or more reverse transcriptases.

2. A composition comprising at least two different SSBs.

9. A method for synthesizing a nucleic acid, said method comprising:

(a) providing a mixture comprising one or more nucleic acid templates, one or anti-DNAP antibodies and/or one or more anti-RT antibodies, and one or more SSBs; and

(b) incubating said mixture under conditions sufficient to synthesize a nucleic acid molecule complementary to all or a portion of said one or more templates.

10. The method of claim 9, wherein said step (a) is accomplished at a temperature sufficient to inhibit nucleic acid synthesis, and wherein said step (b) is accomplished at a temperature sufficient to reduce the inhibitory affect of said one or more antibodies.

14. The method of claim 9 or claim 10, wherein said mixture further comprises at least one component selected from the group consisting of one or more primers, one or more nucleotides, one or more DNAPs, one or more reverse transcriptases, and one or more buffers or buffering salts.

11. A method for synthesizing a nucleic acid, said method comprising:

(a) providing a mixture comprising one or more nucleic acid templates and two or more SSBs; and

(b) incubating said mixture under conditions sufficient to make a nucleic acid complementary to all or a portion of said one or more templates.

12. The method of claim 9 or claim 11, wherein at least one of said one or more templates is cDNA.

13. The method of claim 9 or claim 11, wherein at lest one of said one or more templates is RNA.