Long distance PCR amplification method

Abstract

The invention belongs to the field of molecular biology, and discloses a long-distance PCR amplification method for difficulties such as harsh amplification conditions of longer gene sequences. The amplification method includes the following steps: designing primers, establishing a reaction system, and synthesizing After the primers are added to the established reaction system, the thermal cycle reaction is carried out under certain conditions. After the reaction is completed, the size of the amplified molecule is detected by electrophoresis to confirm that the amplification of the target fragment is completed. The present invention has the advantages of low cost, less materials, etc. advantage.

Description

1. Technical field [0001] The invention belongs to the DNA thermal cycle amplification method in the field of molecular biology. 2. Background technology [0002] Long-distance PCR amplification technology is the basis for further analysis of sequence structural genomics, which can amplify fragments of more than 20kb from the nuclear genome, including isolating complete genes from cDNA (Rose, 1991; Ohler and Rose, 1992; Ohler et al. al., 1993); including the analysis of high molecular weight restriction fragment length polymorphisms in a population (Ohler and Rose, 1992; Ohler et al., 1993). Many confusing questions in life science research will eventually be answered by the start of nuclear genome research. [0003] The advantage of long-distance PCR is that it requires less tissue material, and it can amplify a sufficient number of copies of the target gene DNA sequence from a sample of 1 ng or less (less than 300 copies, which can theoretically b...

AI Technical Summary

Problems solved by technology

Additionally, methodological issues and thermal cycling conditions also affect long-range PCR amplification (Frood and Rose, 1994)