This is the restriction enzyme and acts as “molecular scissors” cuts the two DNA chains at a specific area in the genome so that sections of DNA can be supplemented or detached. A piece of RNA known as guide RNA is the second key molecule. This consists of pre-designed RNA quite small in length sequence, consisting of about 20 bases, positioned within a longer RNA scaffold. The scaffold binds to DNA and the pre-designed sequence ‘guides’ Cas9 to the right part of the genome. ensuring that the Cas9 enzyme intersects at the right point in the genome.
******************************************************************************************** Answer Sheet—Module 7 Lab DNA Extraction Click on the following link and view the DNA extraction: http://learn.genetics.utah.edu/content/labs/extraction/ 1. What is the source of the cells used in this demonstration? A human. 2. Give three practical uses of DNA that is extracted: a.
The restriction enzyme will continue cutting the DNA strand which is broken into fragments that are measured in base pairs (1000 pairs). The restriction enzymes used to analyze DNA in this lab are xHol and Hind III isolated from Xanthomonas holcicola and Haemophilus Influenzae respectively. Plasmids on the other hand used in the lab can be depicted as part of the genetic structure in a cell in which replicate independently on chromosomes, particularly in bacteria consisting of small circular DNA strands. We will construct plasmid maps based on the fragment lengths determined from our results from gel electrophoresis. To prepare our digestion we will combine restriction enzymes along with our plasmid or lambda DNA into 8 microtubes consisting of loading buffer of various proportions measured in microliters.
This complementary base pairing is what makes DNA a suitable molecule for carrying our genetic information—one strand of DNA can act as a template to direct the synthesis of a complementary strand. In this way, the information in a DNA sequence is readily copied and passed on to the next generation of cells. Because of the strict order of the chemical pairing, the double helix design facilitates the correct bonding of the appropriate chemical bases. However, some scientists suggest that the double helix design may also help to increase the physical strength of the gene. Gene construction is anti-parallel, meaning the strands run in opposite directions.
This protein is part of bacterial antiviral defense mechanism and it can destroy double-stranded DNA. Therefore, to prevent lower yield of transfection, we used as host GMO endA negative E coli strain. Plasmid Purification There are several ways to purify plasmids. Plasmid DNA isolation techniques can be simple - low quality DNA preparations “minipreps” and more complex, time-consuming high quality DNA preparations. The high quality DNA preparation requires organic or hazardous cesium chloride ingredients, and the entire process can take a few days.
Enzymes called topoisomerases produce breaks in the DNA molecules and then reconnect the strands, relieving strain and effectively preventing tangling and knotting during replication. DNA polymerase adds new nucleotides to a growing strand of DNA. Because DNA polymerase must adhere to an existing template, an RNA primer is first created at the site of replication. The RNA primer is synthesized by primase, an enzyme that is able to start a new strand of RNA opposite a DNA strand. After a few nucleotides have been added, the primase is displaced by DNA polymerase, which can then add subunits to the 3’ end of the short RNA primer.
Key words: Forensic, DNA, criminal justice system Introduction Forensic science mainly involves the application of Molecular Biology to aid investigation in the criminal justice system. This technology has been facilitated by DNA (Deoxyribonucleic Acid), which is the genetic material of a cell. DNA is the blue print of an organism. Most of the DNA are located in nucleus (nuclear DNA), but a small amount of DNA can be found in mitochondria (mitochondrial DNA). Human DNA consists of about three billion base pairs and more than 99% of those bases are the same in all people.
4-5 A (260 nm)A(280 nm) = 0.3630.175=2.07 For pure RNA: A (260 nm)A (280 nm)≡2.0 ∴ Pellet was almost completely RNA AsA (260 nm)A(280 nm)→1.8, A260 nm can be used to estimate the mass of DNA present Given A260 nm (50 ng µL-1) =1 ∴ m (DNA) = 50 ng X 0.363 = 18.15 ng Volume of solution analysed by U.V. spectrophotometry: 1.0 µL ∴ CDNA = 18.15 ng µL-1 Discussion: E.coli β-galactosidase, encoded for by the lacZ gene, consist of 1024 amino acids [3], therefore lacZ should a minimum codon sequence of about approximately 3.07 kb. BamH1 has one restriction recognition site on the plasmid pRY121 located at the 2780th
DNA EXTRACTION FROM KIWI INTRODUCTION DNA is present in the cells of all living organisms. This procedure is designed to extract DNA from kiwi in sufficient quantity to be seen and spooled. It is based on the use of household equipment and supplies. STUDENT INSTRUCTIONS The process of extracting DNA from a cell is the first step for many laboratory procedures in biotechnology. The scientist must be able to separate DNA from the unwanted substances of the cell gently enough so that the DNA is not broken up.
Each branch uses different kind of technologies as there is availability of technologies which are used in proteomics study. According to Herosimczyk et al., (2006), one of the most effective and widely methods were used are 2-D gel electrophoresis purposely to separate proteins while mass spectrometry purposely to identify and characterize the proteins whereas in expression analysis, protein microarrays are employed in order to study the protein expression. Development of proteomics needs simultaneous advancement of techniques as the challenges is far reaching. Although 2D gel electrophoresis and mass spectrometry play a major role in this study however, there are other technologies which are available which can be used. Figure 1 shows the technologies commonly used which are available and necessary in proteomics study.