Method for preparing DL2000 DNA molecular weight marker as well as product and applications thereof

Abstract

The invention discloses a method for preparing a DL2000 DNA molecular weight marker. The method is characterized by adopting a plasmid enzyme-cut method comprising the following steps of: (1) firstly distributing six fragments to two carriers for amplification, wherein the DL2000 DNA molecular weight marker comprises the six DNA fragments as follows: 100bp, 250bp, 500bp, 750bp, 1kb and 2kb; the carrier 1 comprises the fragment 100bp with the copy number 10, the fragment 250bp with the copy number 4, the fragment 500bp with the copy number 2, the fragment 750bp with the copy number 1, and the fragment 1kb with the copy number 1; and the carrier 2 comprises 2kb with the copy number 1 and the fragment 750bp with the copy number 4; and (2) carrying out enzyme-cutting on the carrier 1 and the carrier 2 by restriction enzyme, and then mixing, thus the molecular weight marker is obtained. According to the invention, the brightness of each belt in the product obtained finally is uniform, the molecular weight marker is convenient to produce, and is convenient for large scale production, and the quality of each batch is stable.

Description

technical field [0001] The invention belongs to the field of bioengineering, in particular to a method for preparing DL2000 DNA molecular weight standards and its products and applications. Background technique [0002] DNA molecular weight standard, commonly known as DNA Marker, is a mixture of a series of DNA fragments of known length, and is the most commonly used reagent in molecular biology experiments. By electrophoresis of the DNA fragment obtained in the experiment and the DNA Marker in the same agarose gel, and by comparing the sample fragment with the DNA Marker, the size of the DNA fragment obtained in the experiment can be roughly estimated. [0003] There are usually two preparation methods for each band of DNA Marker: PCR amplification and plasmid digestion. [0004] The PCR amplification scheme is to design multiple sets of PCR primers for amplification to obtain DNA fragments of corresponding size. Then each fragment was purified and quantified. Finally, t...

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Problems solved by technology

Second, PCR amplification is greatly affected by various conditions, such as instruments, templates, amplification enzymes, different batches of primers, and other reagents, so there will be large differences between different amplification batches

Third, the success rate of PCR amplification is relatively low, and there may be mixed bands in the amplified bands for a long time, or the specificity of the target bands is not very strong, the amplified bands are wider, or the amplified bands may be mixed. Low incremental yields, and other unsuccessful cases

Therefore, there will be large differences between different batches in PCR amplification, and it is very labor-intensive and reagent-intensive, and the production process is difficult to standardize

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