Patent ID: 11873534
Assignee: ELECTROCHAEA GMBH
Field: Biotechnology (Chemistry)
Classification: CPC C  Y | IPC C

Claim 0:
1. A method for detecting the presence of methanogenic microorganism Methanothermobacter thermoautotrophicus DSM3590, comprising:
a. obtaining a sample containing one or more methanogenic microorganisms and releasing DNA from the one or more methanogenic microorganisms to receive an unpurified DNA sample or obtaining a sample containing purified DNA of the one or more methanogenic microorganisms;
b. using the unpurified DNA sample or the purified DNA sample according to step a in a polymerase chain reaction (PCR) amplification using a primer pair comprising a first primer and a second primer, wherein the first primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3; and the second primer comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 4; SEQ ID NO: 5, and SEQ ID NO: 6, wherein each of the first primers and each of the second primers are capable of hybridizing with the DNA of the methanogenic microorganism Methanothermobacter thermoautotrophicus DSM3590 and wherein the primer pair of SEQ ID NO: 1 and SEQ ID NO: 4, or SEQ ID NO: 2 and SEQ ID NO: 5, or SEQ ID NO: 3 and SEQ ID NO: 6 are used to PCR amplify said DNA, to receive an amplified DNA sequence (amplicon), wherein the amplicon comprises at least one restriction enzyme recognition sequence;
c. purifying the amplicon;
d. performing a first digestion reaction with the amplicon of step c by incubating the amplicon in a reaction buffer with at least one first restriction enzyme, which recognizes a first recognition sequence for a time sufficient to form restriction fragments;
e. determining the number of the formed restriction fragments and their size (restriction pattern) and;
f. genotyping the one or more methanogenic microorganisms on the basis of the restriction pattern.