You perform an ELISA to test hospital soap samples for contamination with Pseudomonas aeruginosa. Other than the different molecule of interest, this ELISA works exactly like the one you saw in the video. You run the soap sample, a negative control, and a positive control, all in duplicate. How would you interpret the following results? Laboratory Notebook: ELISA – Pseudomonas aeruginosa Well Contents Observation at 15 min Result 1 Negative control 2 Soap sample 3 Positive control 4 Negative control 5 Soap sample 6 Positive control 7 Empty well 8 Empty well The soap is POSITIVE for the Pseudomonas aeruginosa antigen. You cannot interpret the soap data, because the NEGATIVE control didn’t work. The soap is NEGATIVE for the Pseudomonas aeruginosa antigen. You cannot interpret the soap data, because the POSITIVE control didn’t work.
Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.
Well | Contents | Observation at 15 min | Result |
1 | Negative control | ||
2 | Soap sample | ||
3 | Positive control | ||
4 | Negative control | ||
5 | Soap sample | ||
6 | Positive control | ||
7 | Empty well | ||
8 | Empty well |
The soap is POSITIVE for the Pseudomonas aeruginosa antigen. |
You cannot interpret the soap data, because the NEGATIVE control didn’t work. |
The soap is NEGATIVE for the Pseudomonas aeruginosa antigen. |
You cannot interpret the soap data, because the POSITIVE control didn’t work. |
![The image showcases a series of cuvettes commonly used in spectrophotometric analysis, held in a testing rack. Each of the eight cuvettes is filled with a solution of varying concentrations, indicated by the different intensities of the blue-green color in the fluid inside them. The cuvettes are labeled with the numbers 1 through 8 written in blue marker on their upper parts.
### Detailed Explanation:
1. **Cuvettes and Their Use**:
- Cuvettes are small, rectangular containers used in spectrophotometry to hold samples. They are designed to fit into a spectrophotometer, an instrument that measures how much light a sample absorbs.
2. **Numbering and Concentration**:
- Each cuvette in the image is clearly numbered (1 to 8) with blue ink. The numbers appear to correlate with the concentrations of the solutions they contain. Typically, such numbering would be part of a method to systematically analyze different concentration levels of a substance.
3. **Color intensities**:
- The color variation from light to dark blue-green indicates different concentration levels of a solute in the solvent. Cuvettes 1 and 2 display lighter shades, suggesting lower concentrations, whereas cuvettes 3 through 6 show progressively darker shades, indicative of higher concentrations. Cuvettes 7 and 8 appear to be empty, likely serving as controls or blanks.
4. **Spectrophotometric Analysis**:
- During spectrophotometric analysis, light is passed through these solutions, and the amount of light absorbed by the solution is measured. The absorbance data can then be used to determine the concentration of the solute in each cuvette, following Beer’s Law. This principle states that the absorbance is directly proportional to the concentration of the absorbing species in the solution.
### Educational Context:
Understanding how to prepare and analyze samples using spectrophotometry is crucial in various scientific fields, including biochemistry, molecular biology, and environmental science. The setup shown in the image can be part of a laboratory exercise for students to learn how to correlate the intensity of color (absorbance) with the concentration of a solution.
### Conclusion:
By conducting experiments with cuvettes containing solutions of known concentrations, students can create a standard curve, which is essential for determining the concentration of unknown samples in future experiments. The image exhibits a practical example of setting up such an experiment, emphasizing the importance of careful](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F58a14951-0673-452f-b8e2-b5db778d47b6%2Fa2fa447f-3441-4c1d-9194-5b02ec18bf78%2Fgtx2j7.jpeg&w=3840&q=75)
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