Method for specifically amplifying a dystroglycan, α-sarcoglycan, or endothelin Breceptor cDNA of an extremely small
AssigneeThe Institute of Physical and Chemical Research
InventorsAiji Sakamoto, Fumio Hanaoka
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Abstract
The invention provides a method for selectively amplifying a cDNA synthesized from an mRNA expressed only in an extremely small quantity. The method comprises synthesizing a cDNA from a target mRNA using an oligonucleotide primer complementary to at least a part of the 3′ untranslated region of the mRNA and amplifying the resultant cDNA using a 5′ primer and a 3′ primer which is located 5′ upstream of the oligonucleotide primer for synthesizing a cDNA.
Claims12
A method for detecting a tissue specific cDNA having a coding region which is not actively transcribed in peripheral blood comprising:
- synthesizing the tissue specific cDNA from a target mRNA in the peripheral blood using an oligonucleotide primer shown in SEQ ID NO:
- 1, wherein the tissue specific cDNA encodes dystroglycan and has a base sequence shown in SEQ ID NO:
- 10 or a fragment thereof;
- amplifying the coding region of the resultant cDNA by only one round of PCR using a 5′ primer and a 3′ primer each having 20-40 bases, wherein the 3′ primer is located between the coding region and the oligonucleotide primer;
- and
- detecting the amplified cDNA.
A method for detecting a tissue specific cDNA having a coding region which is not actively transcribed in peripheral blood comprising:
- synthesizing the tissue specific cDNA from a target mRNA in the peripheral blood using an oligonucleotide primer shown in SEQ ID NO:4, wherein the tissue specific cDNA encodes α-sarcoglycan and has a base sequence shown in SEQ ID NO:
- 11 or 12 or a fragment thereof;
- amplifying the coding region of the resultant cDNA by only one round of PCR using a 5′ primer and a 3′ primer each having 20-40 bases, wherein the 3′ primer is located between the coding region and the oligonucleotide primer;
- and
- detecting the amplified cDNA.
A method for detecting a tissue specific cDNA having a coding region which is not actively transcribed in peripheral blood comprising:
- synthesizing the tissue specific cDNA from a target mRNA in the peripheral blood using an oligonucleotide primer sequence shown in SEQ ID NO:7, wherein the tissue specific cDNA encodes endothelin B receptor and has a base sequence shown in SEQ ID NO:
- 13 or a fragment thereof;
- amplifying the coding region of the resultant cDNA by only one round of PCR using a 5′ primer and a 3′ primer each having 20-40 bases, wherein the 3′ primer is located between the coding region and the oligonucleotide primer;
- and
- detecting the amplified cDNA.
A kit for detecting a cDNA encoding dystroglycan in peripheral blood comprising a set of oligonucleotide primers having the base sequence shown in SEQ ID NO:1 for cDNA synthesis and SEQ ID NOS:2 and 3 for PCR.
A kit for detecting a cDNA encoding α-sarcoglycan in peripheral blood comprising a set of oligonucleotide primers having the base sequence shown in SEQ ID NO:4 for cDNA synthesis and SEQ ID NOS:5 and 6 for PCR.
A kit for detecting a cDNA encoding human endothelin B receptor in peripheral blood comprising a set of oligonucleotide primers having the base sequence shown in SEQ ID NO:7 for cDNA synthesis and SEQ ID NOS:8 and 9 for PCR.
Dependencies are inferred from the claim text.