Signal processing method for determining base sequence of nucleic acid

• US 4,958,281 A
• Filed: 10/10/1986
• Issued: 09/18/1990
• Est. Priority Date: 10/11/1985
• Status: Expired due to Term

• Current Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Hara, Makoto
• Primary Examiner(s): Smith, Jerry
• Assistant Examiner(s): BUI, KIM T
• Application Number:
  US06/917,609
• Time in Patent Office: 1,439 Days
• Field of Search: 364/413.01, 364/413.25, 382/6, 935/77, 435/6
• US Class Current: 382/129
• CPC Class Codes: C12Q 1/6869 Methods for sequencing

Abstract

A signal processing method for determining base sequence of nucleic acids by subjecting digital signals to signal processing, said digital signals corresponding to an autoradiograph of plural resolved rows which are formed by resolving a mixture of base-specific DNA fragments or base-specific RNA fragments labeled with a radioactive element in one-dimensional direction on a support medium, which comprises steps of:

(1) preparing at least two one-dimensional waveforms composed of signal position along the resolving direction and signal level for each band;

(2) detecting a position where signal level is maximum on one of the one-dimensional waveforms;

(3) comparing the signal level at said maximum position on the one-dimensional waveform with signal levels at the corresponding positions on one-dimensional waveforms adjacent thereto, to determine a maximum position on the one-dimensional waveform having the highest signal level to be a band position;

(4) detecting a subsequent position where signal level is maximum on said one-dimensional waveform having the band position; and

(5) repeating in order the steps (3) and (4) to thereby determine all band positions on the resolved rows.

Claims

1. A signal processing method for determining base sequence of nucleic acids by subjecting digital signals to signal processing, said digital signals corresponding to an autoradiograph of plural distorted resolved rows which are formed by resolving a mixture of base-specific DNA fragments or base-specific RNA fragments labeled with a radioactive element in a one-dimensional resolving direction on a support medium,

which comprises the steps of:

(1) preparing at least two one-dimensional waveforms composed of signal position along the resolving direction and signal level for each band;

(2) detecting a position where the signal level is maximum on one of the one-dimensional waveforms;

(3) comparing the detected maximum signal level position on the one one-dimensional waveform with signal levels at corresponding positions on adjacent one-dimensional waveforms to determine a maximum position on the one-dimensional waveform having the highest signal level to be a band position;

(4) detecting a subsequent position where the signal level is maximum on said one-dimensional waveform having the band position; and

(5) repeating in order the steps (3) and (4) to thereby determine all band positions on the resolved rows.

2. The signal processing method as claimed in claim 1, wherein said digital signals are obtained by scanning the autoradiograph at a scanning line density that each band is traversed by at least two scanning lines, and in the step (1), said one-dimensional waveforms are prepared for each scanning line.

3. The signal processing method as claimed in claim 1, wherein the mixture of the base-specific DNA fragments consists of the four groups of:

(1) guanine-specific DNA fragments;

(2) adenine-specific DNA fragments;

(3) thymine-specific DNA fragments; and

(4) cytosine-specific DNA fragments;

and the resolved rows consist of four rows formed by resolving each of said four groups of the base-specific DNA fragments on the support medium.

4. The signal processing method as claimed in claim 1, wherein said digital signals corresponding to the autoradiograph are obtained by placing the support medium and a stimulable phosphor sheet comprising a stimulable phosphor together in layers to record the autoradiograph of the plural resolved rows on the phosphor sheet as an energy-stored image, irradiating said phosphor sheet with stimulating rays and photoelectrically detecting the autoradiograph as stimulated emission.

5. The signal processing method as claimed in claim 1, wherein said digital signals corresonding to the autoradiograph are obtained by placing the support medium and a radiosensitive material together in layers to record the autoradiograph of the plural resolved rows on the radiosensitive material as a visible image and photoelectrically reading out the autoradiograph visualized on said radiosensitive material.

6. A signal processing method for determining base sequence of nucleic acids by subjecting digital signals to signal processing, said digital signals corresponding to an autoradiograph of plural distorted resolved rows which are formed by resolving a mixture of base-specific DNA fragements or base-specific RNA fragments labeled with a radioactive element in a one-dimensional resolving direction on a support medium,

which comprises the steps of:

(1) preparing at least two one-dimensional waveforms composed of signal position along the resolving direction and signal level for each band;

(2) detecting positions where the signal level is maximum in corresponding regions on two or more one-dimensional waveforms;

(3) comparing the signal levels at said maximum positions with each other to determine the maximum position having the highest signal level to be a band position;

(4) detecting positions where the signal level is maximum in the corresponding successive region on two or more adjacent one-dimensional waveforms including said one-dimensional waveform having the band position; and

(5) repeating in order the steps (3) and (4) to thereby determine all band positions on the resolved rows.

7. The signal processing method as claimed in claim 6, wherein said digital signals are obtained by scanning the autoradiograph at a scanning line density that each band is traversed by at least two scanning lines, and in the step (1), said one-dimensional waveforms are prepared for each scanning line.

8. The signal processing method as claimed in claim 6, wherein the mixture of the base-specific DNA fragments consists of the four groups of:

(1) guanine-specific DNA fragments;

(2) adenine-specific DNA fragments;

(3) thymine-specific DNA fragments; and

(4) cytosine-specific DNA fragments;

and the resolved rows consist of four rows formed by resolving each of said four groups of the base-specific DNA fragments on the support medium.

9. The signal processing method as claimed in claim 6, wherein said digital signals corresponding to the autoradiograph are obtained by placing the support medium and a stimulable phosphor sheet comprising a stimulable phosphor together in layers to record the autoradiograph of the plural resolved rows on the phosphor sheet as an energy-stored image, irradiating said phosphor sheet with stimulating rays and photoelectrically detecting the autoradiograph as stimulated emission.

10. The signal processing method as claimed in claim 6, wherein said digital signals corresponding to the autoradiograph are obtained by placing the support medium and a radiosensitive material together in layers to record the autoradiograph of the plural resolved rows on the radiosensitive material a a visible image and photoelectrically reading out the autoradiograph visualized on said radiosensitive material.