The invention relates to an ISSR(Inter-simple
Sequence Repeat)-PCR(
Polymerase Chain Reaction)
molecular marker method for macromitrium gymnostomum. The ISSR-PCR
molecular marker method comprises the following steps of firstly, extracting
genome DNA (
Deoxyribose Nucleic Acid) of the macromitrium gymnostomum; secondly, performing ISSR-PCR amplification, wherein a
reaction system for performing the ISSR-PCR amplification in step (2) is as follows: 30mu L
reaction system contains 3mu L of 10*LAPCR (
Polymerase Chain Reaction )
buffer solution, 50ng of template
DNA, 1.5mmol / L of Mg<2+>, 0.25mmol / L of dNTP (
Deoxyribonucleoside Triphosphate), 1.5U of Taq
enzyme and 0.5mu mol / L of primer. A primer UBC(University of British Columbia) 834 is taken as an example, ISSR amplification comprises conditions of: annealing at the temperature of 94DEG C for 4 minutes, then annealing at the temperature of 94DEG C for 45 seconds, at the temperature of 53DEG C for 45 seconds and at the temperature of 72DEG C for 1 minute for 45 circles, and finally, prolonging the annealing time at the temperature of 72DEG C by 10 minutes. Compared with the prior art, the ISSR-PCR
molecular marker method disclosed by the invention has the advantages that a strip amplified by an established ISSR-PCR
reaction system for the macromitrium gymnostomum has the characteristics of high stability,
high definition and high polymorphism; the shortages in the research on the
genetic diversity of the macromitrium gymnostomum are made up; the achievements of the invention can be used for researching the genetic relationship, a molecular marker and
biogeography, and has high scientific value and application value.