Restriction Fragment Length Polymorphism Lab Report
Restriction Fragment Length Polymorphism and Southern Blotting
1. Abstract
The aim of the experiment was to be introduced to the techniques involved in the identification of restriction fragment length polymorphisms. Restrictions were carried out using three different restriction enzymes, ECORI, HindIII and BstELI with their buffers. Lambda (λ) DNA was then examined using electrophoresis and Southern blotting.
The results showed that λ DNA was best digested by EcoRI as all of the expected bands can be seen and are in the right place. The HindIII digestion was second best as there was a large amount of merging and the bands contained a lot more streaking compared to the other enzymes. The BstELI was only partially digested. Many bands had not separated, and the other bands were either in the wrong location or barely visible.
2. Introduction
2.1 RFLP
Restriction fragment length polymorphisms (RFLPs), were developed by Botstein et al. in 1980. They are differences in the lengths of DNA fragments cut by restriction enzymes at specific sequences called restriction sites. …show more content…
Dr. Alec Jeffreys used RFLP to analyse DNA and discovered that repetitive patterns of DNA were present in all human beings, but they were different in length for all individuals. He realised that the variation could be used to ascertain the identity of a person and called his technique genetic fingerprinting. The process is used as a means of identification when an assailant has left some kind of bodily fluid at the crime scene but no visual identification is possible. Genetic fingerprinting relies on the principle that apart from identical twins no other individuals share the same genetic code. Jeffreys' technique was catapulted into the forensic science domain when two murders were committed and genetic fingerprinting was used to exonerate a suspect and convict the guilty
1. Abstract
The aim of the experiment was to be introduced to the techniques involved in the identification of restriction fragment length polymorphisms. Restrictions were carried out using three different restriction enzymes, ECORI, HindIII and BstELI with their buffers. Lambda (λ) DNA was then examined using electrophoresis and Southern blotting.
The results showed that λ DNA was best digested by EcoRI as all of the expected bands can be seen and are in the right place. The HindIII digestion was second best as there was a large amount of merging and the bands contained a lot more streaking compared to the other enzymes. The BstELI was only partially digested. Many bands had not separated, and the other bands were either in the wrong location or barely visible.
2. Introduction
2.1 RFLP
Restriction fragment length polymorphisms (RFLPs), were developed by Botstein et al. in 1980. They are differences in the lengths of DNA fragments cut by restriction enzymes at specific sequences called restriction sites. …show more content…
Dr. Alec Jeffreys used RFLP to analyse DNA and discovered that repetitive patterns of DNA were present in all human beings, but they were different in length for all individuals. He realised that the variation could be used to ascertain the identity of a person and called his technique genetic fingerprinting. The process is used as a means of identification when an assailant has left some kind of bodily fluid at the crime scene but no visual identification is possible. Genetic fingerprinting relies on the principle that apart from identical twins no other individuals share the same genetic code. Jeffreys' technique was catapulted into the forensic science domain when two murders were committed and genetic fingerprinting was used to exonerate a suspect and convict the guilty