197 results about "Genome amplification" patented technology

The invention relates to a marker primer interlocked with a wheat powdery mildew resistance gene PmHNK54 and application thereof in molecular marker-assisted selection. The sequence of the marker primer on a genetic map is Xbarc5, PnHNK54 and Xgwm312, and the genetic distances between Xbarc5 and PnHNK54 and between Xgwm312 and PnHNK54 are respectively 5.0cM and 6.0cM. 10 mu l of PCR (Polymerase Chain Reaction) reaction system during SSR (Simple Sequence Repeat) marker selection comprises 1 mu l of 10*buffer, 0.2 mu L of 10nM / mu l dNTP, 0.2 mu l of 20u M / mu L primer, 0.1 mu l of 5U / mu lTaq enzyme, 8.1 mu l of deionized water and 0.4 mu l of 20 ng / mu l genome DNA. An amplification procedure comprises the following steps of: pre-denaturing at 94 DEG C for 3 min, denaturing at 94 DEG C for 1 min, renaturing at 55 or 60 DEG C for 1 min, extending at 72 DEG C for 2 min, and repeating for 40 cycles; extending at 72 DEG C for 10 min; and performing 6% denaturing polyacrylamide gel electrophoresis or 8% nondenaturing polyacrylamide gel electrophoresis on amplification products, observing and taking pictures after silver nitrate dyeing. The invention finds the position of the powdery mildew resistance gene PmHNK54 in wheat parent material Zheng 9754, has the advantages of strong marker specificity, high stability, cost saving and high selection efficiency and is applicable to large scale, high throughput and automation.

A method for whole genome amplification comprising (a) treating genomic DNA with a modifying agent which modifies cytosine bases but does not modify 5′-methyl-cytosine bases under conditions to form single stranded modified DNA; (b) providing a population of random X-mers of exonuclease-resistant primers capable of binding to at least one strand of the modified DNA, wherein X is an integer 3 or greater; (c) providing polymerase capable of amplifying double stranded DNA, together with nucleotides and optionally any suitable buffers or diluents to the modified DNA; and (d) allowing the polymerase to amplify the modified DNA.

The invention discloses a spider mitochondrial genome sequence amplification primer and amplification method, including the extraction of the spider genome total DNA, the design of a universal amplification primer for the mitochondrial genome sequence, and the long PCR (L-PCR) of the mitochondrial genome DNA Amplification and sequencing. The primers for the amplification of the whole mitochondrial genome sequence of spiders consisted of 6 pairs of single-stranded oligonucleotides. The universal amplification primer of the present invention can rapidly and efficiently amplify the full sequence of mitochondrial genes of various spiders, laying a material foundation for the identification of spider species, population diversity analysis, germplasm resource protection and systematic genetic evolution research.

Disclosed are methods and compositions for amplification of genetic material, including isothermal WGA of single cells.