Gel Electrophoresis Lab

This is a sheet of special blotting paper. The DNA fragments keep the same pattern of separation that they had on the gel. The blot is incubated with multiple copies of a probe. The probe forms base pairs with the complementary the DNA sequence and binds to form a double stranded DNA molecule. This step is called hybridisation.

n.d.). DNA samples are submitted to a certified laboratory and undergo the following process (DNA Evidence. n.d.): • Extraction is the process of releasing the DNA from the cell. • Quantitation is the process of determining how much DNA you have. • Amplification is the process of producing multiple copies of the DNA in order to characterize it.

Rosalind discovered that DNA could exist in two forms and also discovered that within her x-Ray of DNA, the wet form of DNA had all the characteristics of a helix. Watson and Crick, after later looking at Franklin’s results, suggested that the molecules of DNA were made of two

PRACTICAL 4 Materials and Methods Measurement of DNA concentration The most common technique to measure DNA concentration is measurement of absorbance. We had used 1:20 dilution of the DNA sample and the reading was expected to be in the range of 0.1-1.5 OD260. 5µl of DNA and 95µl of PBS buffer were mixed together and inserted into a clean cuvette. Then it was put inside the spectrophotometer. The measurement was taken at 230nm, 260nm and 280nm.

Since the SDS is negatively charged, sample proteins move to the positively charged anode through the gel. Small proteins migrate faster since it is light and is seen further below the gel than the larger proteins which migrate slowly and stays at the top of the gel. Therefore, proteins are separated according to their size in gel electrophoresis. A notch is made at the top

They mostly involve sophisticated machinery and staining techniques that have high-throughput results. A. QF-PCR: Quantitate Fluorescent Polymerase Chain Reaction involves detection of chromosome specific DNA sequences known as genetic markers or short tandem repeats (STRs). It involves the use of primers labeled with fluorescent tags for PCR amplification of individual markers and the copy number of each marker is indicative of the copy number of the chromosome. The resulting PCR products may be analyzed and quantified using an automated genetic analyzer. The genetic markers may vary in length across individual patients and chromosomes, depending on the no.

23 inherited from the father called paternal and 23 from the mother called maternal. On the 46 chromosomes there are alleles that code for certain traits. If some allelic traits are not expressed (imprinted) can cause genetic disorders like Prader-Willi or Angelman syndrome (2013, Klug). Prader-Willi Syndrome and Angelman Syndrome Prader-Will and Angelman are two

Potassium ions diffuse out the cell due to the concentration gradient, creating a potential difference across the membrane. Other ions, such as sodium, are unable to cross the membrane and thus remain concentrated on one side. Consequently, the increased negative charge created inside the cell attracts potassium ions back across the membrane into the cell. This force is called electrostatic pressure. When the potential difference across the membrane is around -70mV, the electrical gradient exactly balances the chemical gradient and equilibrium is reached.

Factors affecting the double-helical structure of DNA Besides having hydrogen bonds between the bases to hold the two DNA strands together, the backbone of the polynucleotides must be highly charged too. A strong repulsion between the two strands and will cause them fall apart if there is no salt in the surrounding medium. Thus counter-ions in the surroundings are essential for the double helical structure as they serve as shields for the charges on the sugar-phosphate backbone. These ions also provides attractive interactions with the fluctuating counter-ions around the backbone for fluctuating induced dipoles. Under different conditions such as hydration, thermal fluctuations or applied force and/or twist, the DNA adopts several different helical

Pre-Lab Questions 1. Compare and contrast mitosis and meiosis. Mitosis and Meiosis both include splitting DNA between new cells. They both include cell reproduction which contains chromosomes from both parents. In Mitosis the two daughter cells are identical from a single parent cell.

The purpose of this experiment is to create a complete genomic library of Aliivibrio fisheri through the use of the lux operon. The examination of the lux operon gene occurs through the extraction of the DNA of Aliivibrio fischeri and digest a large piece of DNA to smaller random pieces. The fragment of DNA will later be ligated together in plasmid. Plasmid acts as vectors to transport DNA from one organism to another. The DNA will then run through a UV-visible spectrophotometer to test the absorbance of the extracted DNA.

The parents turn out different based on DNA combos that are chosen. 4. What is the term for the random arrangement of homologous pairs of chromosomes during the first division of meiosis? Independent Assortment 5. What role does the Polymerase Chain Reaction (PCR) play in producing a DNA Profile?

A Gly-chloride ion boundary is formed since glycine moves slower than chloride ion. However, glycine still runs slightly faster than other proteins. As a result, the glycine keeps pushing the protein towards the chloride ion. In other words, the proteins are trapped between glycine and chloride ion. The proteins form a very tight band inside the stacking gel.

To begin the process, a pre-prepared 1% agarose gel was obtained. The gel chamber was set up by placing the agarose gel into the chamber and submerging it in plentiful TAE buffer. The wells were filled with both PCR and DNA and shared between six students. The wells were labeled 1-7. The first well was pre-loaded with DNA ladder and labeled as 1 microliter kb DNA ladder.