Method for detecting forest chromosome end by using oligo sequence

Abstract

The invention provides a method for detecting the forest chromosome end by using an oligo sequence. The method is implemented by an (AGGGTTT)3 oligo sequence probe and a kit comprising the probe, thebase sequence of the (AGGGTTT)3 oligo sequence probe is: 5'-AGGGTTTAGGGTTTAGGGTTT-3',and the probe is used for identifying and labeling the forest chromosome end; the kit comprises the probe, an enzymatic hydrolysate, a 20*SSC buffer liquor, a 2*SSC buffer liquor, a 70% FA liquor, a 50% DS liquor and salmon sperm DNA. The method comprises the following steps: obtaining of a forest chromosome slidespecimen; denaturing; FISH (fluorescence in-situ hybridization); signal detection; slide recovery. According to the method, a slide can be used repeatedly, the probe is easy and quick to prepare andhigh in sensitivity. The probe is only directed at the oligo sequence at the chromosome end, the root slide is easy to make, the metaphase chromosome is convenient to obtain, and the whole experimental process requires short time and is easy to operate.

Description

technical field [0001] The invention belongs to the application field of fluorescent in situ hybridization probes, and particularly relates to a method for detecting the ends of forest tree chromosomes by using oligosequences. Background technique [0002] At present, most of the methods for detecting forest tree chromosomes are simple microscopic observation, taking photos and analyzing them. Compared with herbaceous plants, forest tree chromosomes are smaller. In many cases, the number, size and shape of chromosomes cannot be clearly distinguished, resulting in a huge experiment. error. [0003] Fluorescence in situ hybridization (FISH) is a non-radioactive molecular cytogenetic technology developed on the basis of radioactive in situ hybridization. method. FISH can observe forest tree chromosomes more intuitively, efficiently and quickly, but fluorescent labeling of chromosomes has become one of the difficulties. In addition to the lack of a simple and fast method for ...

AI Technical Summary

Problems solved by technology

FISH can observe forest tree chromosomes more intuitively, which is efficient and fast, but fluorescent labeling of chromosomes has become a difficulty

In addition to the lack of a simple and fast method for fluorescent labeling of chromosomes, there are also problems such as the large amount of target DNA required in the labeling process, which may cause labeling errors, repeated labeling, and incomplete and unclear chromosome numbers.

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