Increased throughput analysis of small compounds using multiple temporally spaced injections

• US 6,533,912 B2
• Filed: 05/16/2001
• Issued: 03/18/2003
• Est. Priority Date: 07/13/1999
• Status: Expired due to Term

• Current Assignee: IntegenX Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Molecular Dynamics Inc. (General Electric Company)
• Inventors: Peponnet, Christine, Mansfield, Elaine S., Bashkin, John S., Kautzer, Curtis R.
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): Kim, Young
• Application Number:
  US09/859,840
• Publication Number:
  US 20010053554A1
• Time in Patent Office: 671 Days
• Field of Search: 204/457, 204/450, 204/453, 204/456, 204/604, 356/344, 435/6, 435/91.2
• US Class Current: 204/457
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00587 High throughput processes
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44782 of a plurality of samples

Abstract

The apparatus and method of the present invention disclose a system in which multiple injections may be made into a capillary array. The injections are spaced in time with each injection followed by an interval of electrophoresis. Once all samples are loaded into the capillaries, continuous electrophoresis and detection is used to separate and detect target compounds within the sample. The interval between injections is matched to the target compound migration rate to be sufficient to allow the target compounds to be detectably separated when the compounds reach the detector.

Claims

1. A method of analysis of molecules in a capillary dimension channel(s) filled with a separation media, the method using electrophoretic separation to separate small compounds, the method comprising:

a) injecting a sample into a injection end of said capillary dimension channel, said sample including at least one small molecule and at least one quality metric standard compound;

b) introducing an electric field through the capillary dimension channel for a selected period of time, whereby said electric field causes compounds in said sample to migrate away from said injection end;

c) repeating steps a and b at least one time; and

d) detecting at a detection location said at least one small molecule and at least one quality metric standard compound;

wherein said detection location is separated from said injection end by a length of the capillary dimension channel;

wherein said selected period of time is sufficient to allow the compounds in each sample to be detectably separated at the detection location;

wherein every sample injected into the capillary dimension channel is continuously detected.

2. The method of claim 1, wherein the step of injecting said sample includes injecting a sample containing nucleic acids.

3. The method of claim 1, wherein the step of injecting said sample includes injecting a sample containing a DNA fragment.

4. The method of claim 1, wherein the step of injecting said sample including at least one quality metric standard includes injecting a sample including a nucleic acid quality metric standard.

5. The method of claim 1, wherein the method is effected in a plurality of capillary dimension channels in an array of capillary dimension channels.

6. The method of claim 1, wherein the step of detecting is effected by illuminating the detection location and detecting a detection signal produced by the small molecule and the quality metric standard compound, said detection signal excited by illuminating the detection location.

7. The method of claim 6, wherein the step of illuminating the detection location includes illuminating using laser illumination.