Nucleic acid marker ladder for estimating mass

• US 5,834,201 A
• Filed: 07/11/1997
• Issued: 11/10/1998
• Est. Priority Date: 10/28/1993
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Incorporated (Thermo Fisher Scientific Incorporated)
• Inventors: Hartley, James L.
• Primary Examiner(s): Houtteman, Scott W.
• Application Number:
  US08/893,523
• Time in Patent Office: 487 Days
• Field of Search: 435/6, 435/91.2
• US Class Current: 435/6.12
• 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: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]

Abstract

The invention relates to a nucleic acid marker ladder which is a restriction endonuclease digest, wherein a nucleic acid restriction endonuclease digest is a collection of nucleic acid fragments resulting from complete digestion of one or more nucleic acids by one or more restriction endonucleases; the restriction endonuclease digest contains at least 3 fragments; and the size of the fragments in base pairs is a multiple of an integer, wherein the integer is 10 or more.

Claims

1. A nucleic acid marker ladder consisting essentially of a restriction endonuclease digest, wherein

(a) the nucleic acid restriction endonuclease digest is a collection of nucleic acid fragments resulting from the complete digestion of one or more nucleic acids by one or more restriction endonucleases;

(b) the restriction endonuclease digest contains at least 3 fragments; and

(c) the size of the fragments in base pairs is a multiple of an integer, wherein the integer is 10 or more.

2. The nucleic acid marker ladder according to claim 1, wherein the integer is 10.

3. The nucleic acid marker ladder according to claim 1, wherein the integer is 25.

4. The nucleic acid marker ladder according to claim 1, wherein the integer is 50.

5. The nucleic acid marker ladder according to claim 1, wherein the integer is 100.

6. The nucleic acid marker ladder according to claim 1, wherein the collection of nucleic acid fragments results from digestion of a nucleic acid by one restriction endonuclease.

14. A method of using a nucleic acid marker ladder to estimate the mass of a nucleic acid comprising:

(a) electrophoresing a known amount of the marker ladder of claim 1 and an unknown amount of said nucleic acid on an agarose gel; and

(b) comparing the mass of said marker ladder with the mass of said nucleic acid.

15. The nucleic acid marker ladder according to claim 1, wherein in (a) the collection of nucleic acid fragments is generated simultaneously in one reaction.

7. A nucleic acid marker kit comprising a carrier means having in close confinement therein at least one container means where the first container means contains a nucleic acid marker ladder consisting essentially of a restriction endonuclease digest, wherein

(a) the nucleic acid restriction endonuclease digest is a collection of nucleic acid fragments resulting from the complete digestion of one or more nucleic acids by one or more restriction endonucleases;

(b) the restriction endonuclease digest contains at least 3 fragments; and

(c) the size of the fragments in base pairs is a multiple of an integer, wherein the integer is 10 or more.

8. The nucleic acid marker kit according to claim 7, wherein the integer is 10.

9. The nucleic acid marker kit according to claim 7, wherein the integer is 25.

10. The nucleic acid marker kit according to claim 7, wherein the integer is 50.

11. The nucleic acid marker kit according to claim 7, wherein the integer is 100.

12. The nucleic acid marker kit according to claim 7, wherein the collection of nucleic acid fragments results from digestion of a nucleic acid by one restriction endonuclease.

13. A method of preparing a nucleic acid marker ladder comprising:

(a) generating at least two polymerase chain reaction (PCR) products wherein each product is generated from a template comprising a restriction endonuclease site and a primer comprising the restriction endonuclease site in the template;

(b) joining the PCR products to produce a nucleic acid molecule; and

(c) completely digesting one or more nucleic acid molecules with at least one restriction endonuclease

wherein a nucleic acid marker ladder is produced wherein the ladder contains at least 3 fragments and the size of the fragments in base pairs is a multiple of an integer, wherein the integer is 10 or more.

16. The method of claim 13, wherein the nucleic acid marker ladder is prepared by the complete digestion of one or more nucleic acids with one restriction endonuclease in step (c).

17. The method of claim 13, wherein the fragments comprising the marker ladder are prepared simultaneously in one reaction in step (c).