Cottonrose hibiscus ISSR-PCR reaction system, molecular marking method and application

• Main Authors: LI FANGWEN, TANG SHENGWEN, ZENG XINMEI, ZHU ZHANGSHUN, SHI XIAOQING, MA JIAO, CHEN XI
• Format: Patent
• Language: Chinese; English
• Published: 18.06.2024
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; NEW PLANTS OR PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TRAPPING; VINEGAR; WINE

The invention relates to a cottonrose hibiscus ISSR-PCR reaction system and application. Every 25 [mu] L of the reaction system contains 50 ng of template DNA, 3.5 [mu] l of Easy Taq Buffer (with Mg < 2 + >), 1.5 [mu] l of a primer, 0.2 [mu] l of Taq polymerase and 0.8 [mu] l of dNTPs. And amplifying the reaction system according to the following procedures: pre-denaturation at 95 DEG C for 5 minutes, denaturation at 94 DEG C for 1 minute, annealing of each primer at the optimal annealing temperature for 1 minute, extension at 72 DEG C for 2 minutes, circulation for 40 times, supplementary extension at 72 DEG C for 10 minutes, and preservation at 4 DEG C. The invention also provides an ISSR-PCR molecular marking method for cottonrose hibiscus. A stripe amplified by the established hibiscus mutabilis ISSR-PCR amplification reaction system is high in definition and stability, has very high polymorphism, and makes up for the deficiency of the current research on the genetic diversity of hibiscus mutabilis; the h