Materials and Methods Restriction enzyme digestion is a method used to identify any differences in DNA sequences among people, the use of restriction enzymes are what help to determine the order of DNA in which they attach to recognition sites and cut the DNA. People with different genes have different recognition sites for restriction enzymes. (Upadhyaya 45-46) In this experiment, four test tubes were used for restriction enzyme digestion reactions which were all filled using a micropipette with
The first step in genetically modifying cotton is to remove the gene Cry1Ac from the bacteria Bacillus thuringiensis using a specific restriction enzyme. A restriction enzyme cuts DNA strands at highly specific sites, diagonally, leaving some of the nucleotides from each strand exposed on either end of the required gene. These are called ‘sticky ends’, making it easier to attach the gene into another piece of DNA. The second step is to prepare a bacterium (agrobacterium tumerfaciens) to carry the
in life. The process of genetically modifying babies is quite complex and there are many steps involved in this process. The basic process of this is, a embryo will be created through IVF (the mothers egg and fathers sperm will be fertilised in the lab). Once fertilisation has been completed a cell will be removed from the embryo and tested to identify any diseases. If the embryo is healthy then it can be inplaned into the mother’s womb, however if the test indicates that the embryo has a disease
PCR products were run on 1 % agarose gel. To verify the transformation of the clones, restriction enzyme digestion was employed to cleave three different plasmids: pGEX-GFPS65T-1, - 2 and - 8, at specific sites. This was done with following restriction enzymes (RE): BamHI + PstI and EcoRI + PstI. A 1% agarose gel was run on the resultant fragments. The procedure for the experiment is outlined in details in the lab manual Laboratory Notes BIOC6017 Part1 (p.16-21). 2.2 Purification of recombinant GST-GFP
cells. This is done by using the sugar arabinose. When the araC protein binds to the DNA sequence and is turned on in the presence of arabinose and activates it and it helps the araC bind to the RNA polymerase and transcribes the araA, araB, and araD enzymes. The coding sequence of the pGLO has been modified to include many of the inner workings of the arabinose operon which uses arabinose as an inducer. The promoter (PBAD) and the araC gene are present in the process of translation. The genes that code
blocks the binding of RNA polymerase which will inhibit the process of transcription for the utilisation of glucose. Lip genes and the repressor blocks the binding of primase which is involved in providing a starting point for DNA polymerase. If this enzyme is inhibited, the synthesis of the new DNA strand will be halted. It is also designed for the utilisation of lactose that can also be lacl regulatory genes and the repressor blocks the movement of helicase which leads to the DNA strands being unable
1. What is the principle of polymerase chain reaction? Polymerase chain reaction (PCR) is a process to amplify the gene once the proper gene of interest has been cut using the restriction enzymes . This is accomplished by specific proteins known as polymerases, enzymes that are able to string together individual DNA building blocks to form long molecular strands. Through this process, multiple copies of the gene of interest can be produced. PCR proceeds in three stages, denaturation, annealing
APPLICATIONS OF CELL CULTURE Cell culture is one of the major techniques used nowadays in many applications, since it has a significant role in research, production and improvement of many products. Cell culture provides model systems being used for the study of the physiology and basic biochemistry of cells such as aging and cellular metabolism, for the study of effects of new drugs on cells, and to study mutagenesis as well as carcinogenesis. Animal cell culture technique can be used to study
BIOFUELS Two problems roused the world’s concern in recent years. One is energy crisis caused by the squandering of petroleum fuel. Second problem is environmental problems such as greenhouse effect, global warming, etc. To deal with such problems, renewable sources, utilization technology and bioenergy production technology have been developed. Bioethanol as one of the biofuel can be blended in different proportions in automobiles with gasoline. Biobutanol is also a new biofuel (Liu, Wang, & Zhang