The proteins can then be removed from the antibodies and separated using gel electrophoresis techniques. A very common technique often used is called two-dimensional gel electrophoresis. A sample is run on a very thin strip of polyacrylamide then placed under a perpendicular current, moving the proteins within the sample first in one direction, then separating them in another. This allows separation of molecules by size and by differing charge of molecules of similar molecular weights. Most useful to the fields of biochemistry and molecular genetics is the use of these methods in gene identification.
In addition, diseases such as Huntington’s disease, breast cancer, and muscular dystrophy are presently being screened for in humans (Jaroff, 1996). How researchers are able to screen for genes New developments have given researchers the ability to decipher the genetic code of organisms. Some of the techniques that researchers use are RFLP (restriction fragment length polymorphism) analysis and DNA probes. RFLP analysis utilizes enzymes from bacteria that are thought to be used as defense mechanisms against invading viral DNA. The enzymes fragment foreign DNA at specific locations depending on the base sequence (Griffiths, 1996).
According to Yourgenome.org, Gene therapy is basically when the DNA is introduced into a patient to treat a genetic disease. Precisely, the new DNA contains a functioning gene typically to correct effects of a disease-causing mutation. Typically, there are two types of gene therapy, but they are different depending on the kinds of cells treated. There is somatic gene therapy and germline gene therapy. Additionally, there are also some techniques used to carry out gene therapy.
Gel electrophoresis is a procedure used in laboratories to separate DNA, as well as RNA and proteins. A gel slab is placed in a buffer-filled box and an electrical field is applied. The negatively charged DNA will migrate towards the positively charged side, where it can then be recorded and further analyzed. An example for the use of gel electrophoresis would be in identifying people. DNA is present in almost every cell of our body.
Restriction fragment length polymorphism (RFLP) analysis is a technique in which DNA regions are digested using restriction endonucleases, and subjected to radioactive complementary DNA probes to compare the differences in DNA fragment lengths between individuals. The DNA in question is digested using restriction endonuclease(s). The DNA is then run on a gel and appears to be very long. The gel is subject to a chemical that causes the double-stranded DNA to separate into to individual strands. The strands are then transferred to a nylon membrane with using an electric current, where it will bind.
The polymerase chain reaction or PCR for short can be used to create many copies of DNA. This allows the DNA to then be visualized using a dye like ethidium bromide after gel electrophoresis. The process has been refined over the years, however the basic steps are similar. The first is to denature dsDNA through heating to ~96 °C. This separates the two strands of DNA.
There are several applications for genetic engineering in microbiology as well as other fields of biology.It includes invitro mutagenesis,gene synthesis, Expressing eukaryotic genes in bacteria,production of transgenic plants and animals,gene therapy,screening for genetic diseases and forensic analysis. STEPS IN RECOMBINANT DNA TECHNOLOGY 1) PURIFYING DNA/ISOLATION OF DNA Purifying DNA and isolating genes is an important step in geneticengine... ... middle of paper ... ...e for the synthesis of new DNA strands, each randomly terminating due to the incorporation of a chain terminating dideoxynucleotide in 4 different reaction tubes. This produces a population of molecules, each terminating at a different site. Running the products in each tube on a gel allows the determination of where each chain terminating dideoxynucleotide was incorporated. The DNA is visualized because the DNA primer to start the reaction is radioactive or some of the dNTPs are radioactive This procedure is now automated so that a computer reads the sequence.
Another molecule of DNA that had also been snipped with the same restriction enzyme was found to have a corresponding sticky end that could combine with the original sticky end to form what scientists call a recombinant DNA molecule. By using restriction enzymes, scientists can cut genes out of chromosomes in order to reinsert them into other ... ... middle of paper ... ...eld of medicine. Both genetic engineering and stem cell technology are essential branches of biotechnology. Genetic engineering is the skillful manipulation of a gene by way of a process other than ordinary reproduction. Stem cells that won’t be denied by immune systems, can divide almost forever, and can alter themselves easily into other cells are used in stem cell technology and hold much promise in the cure of diseases and injuries.
What is important when you design primers? there are several considerations for primers design including primer length between 18 and 24 bases, Melting Temperature (Tm) , 3' sequence and guanine-cytosine content .  3. Describe how PCR is performed! ● step 1- DNA , tow primer (forward and reverse primer), dNTPs , and DNA polymerase enzyme mixed together in sutable tube.
The temperature at which the two complementary strands separate is called the melting temperature ‘Tm’, and is affected by the percentage of G.C base pairs, ion concentration of the solution, presence of destabilising compounds like urea, and the pH of the solution (Lodish et al, 2004, p. 105). Nucleic acid... ... middle of paper ... ...ected to a probe. These techniques can be used to distinguish between alleles that vary even by single nucleotides (“Nucleic acid hybridization assays”, 1999, Ch. 5). Nucleic acid hybridisation is used in many routine experiments in the molecular biology laboratory, making it an indispensable requirement in genetic engineering and molecular biology.