Using PCR and Gel Electrophoresis to Determine Genotype

Determining genotype using PCR and gel electrophoresis In some cases it is necessary to identify the DNA recovered from the sample. When a small sample to be identified is required, the DNA in the sample is multiplied by multiple identical samples using the polymerase chain reaction. The DNA recovered from the reaction can then be introduced into apalatas using gel electrophoresis and the DNA sample can be compared with other samples. In our experiment we learned how to copy small DNA samples to usable amounts and how to analyze specimines using gel electrophoresis.

Denaturing gradient gel electrophoresis (DGGE) is a method for isolating PCR product DNA products for molecular fingerprinting. DGGE differs from conventional agarose gel electrophoresis in that it separates PCR products based on sequence size and their rate of denaturation. In contrast, traditional agarose gel electrophoresis is based solely on size separation. This limitation makes agarose gel electrophoresis impossible for molecular fingerprinting methods. The reason is that because some samples may have similar size PCR fragments, only one or more slightly spaced bands are formed and it is not possible to determine It will be possible.

Isolation of the resulting PCR amplicons by PCR amplification, denaturing gradient gel electrophoresis (DGGE) or temperature gradient gel electrophoresis (TGGE) of the 16S rRNA gene (16S rDNA) using consensus bacterial primers and cloning was used Constitute the most common molecule. Ecological techniques describe soil bacterial ecology to date (42). Clones or bands on the gradient gel can be sequenced and the information obtained can be used to infer the diversity of the original sample. Over the past few years, these studies have been increasingly applied in soil (14, 29, 47). Because molecular technology has been systematically applied in many different environments. To date, the greatest contribution of these studies to soil microbiology would be a sequence-based taxonomy. Based on the ribosomal sequence, we observed a dramatic change in the bacteria "kingdom" and evolutionary method (26)