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Determining nucleotide sequence of a gene to determine amino acid sequence of a protein, gene and protein structure, identify regulation sequences and mutations and differences in genes from splicing

Dideoxyribonucleotide (ddNTP) is mixed with a vector, primer, polymerase, and deoxyribonucleotides, and ddNTP incorporates into DNA to prevent extension

In PCR, primer has DNA pol start DNA synthesis, and randomly insert ddNTP into DNA to prevent further synthesis, and create fragments in the DNA

Bands are later separated on polyacrylamide gel to differ in length by single nucleotides, and autoradiography detects radioactive sequences

Original experiment by Fredrick Sagar could identify sequences up to 200-400 nucleotides, by computer-automated programs can detect up to 500 nucleotides per reaction

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Determining where a gene is in the DNA by mapping where the restriction enzymes cut the gene out after the gene has been cloned

Useful for making small copies of the gene and manipulating small pieces of DNA to sequence DNA and use probes to study expression

Done by first digesting the DNA with restriction enzymes, then doing double digestions with combined enzymes, fragments are separated by electrophoresis, then single and double digests are compared to each other to order them into a restriction map

Done today with bioinformatics software

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is a cytogenetic technique used to detect the presence or absence and location of specific gene sequences.

It can visualize specific cytogenetic abnormalities (copy number aberrations) such as chromosom-al deletion, amplification, and translocation.

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Fluorescence In Situ Hybridization (FISH) is used in different areas:

1. Medical Testing: It helps diagnose genetic diseases like Down syndrome and various cancers.

2. Cancer Research: FISH detects genetic changes in cancer cells, aiding research and patient care.

3. Prenatal Screening: It checks for genetic issues in unborn babies during pregnancy.

4. Microbiology: FISH identifies bacteria, viruses, and fungi for research or diagnosing infections.

5. Plant Genetics: It helps study plant genes and breeding for better crops.

6. Forensics: FISH assists in DNA analysis for criminal investigations and paternity testing.

7. Developmental Biology: It studies gene patterns during embryo growth and tissue formation.

8. Environmental Monitoring: FISH assesses microorganisms in ecosystems, aiding environmental studies.

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Fluorescence In Situ Hybridization (FISH) was developed by biomedical researchers Joseph Gall and Mary Lou Pardue in the early 1980s. They published their groundbreaking work on FISH in 1981, detailing its application for visualizing specific DNA sequences within chromosomes. Since then, FISH has become a widely used technique in molecular biology, genetics, and various other fields.

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DNA microarray is a tool used to determine whether the DNA from a particular individual contains a mutation in genes.

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Gene microarrays are used in various fields of biological and medical research, as well as in clinical diagnostics. Some common applications include:

1. Gene expression profiling: Microarrays are used to measure the expression levels of thousands of genes simultaneously. This allows researchers to study how gene expression patterns change in response to different conditions or stimuli, providing insights into biological processes, disease mechanisms, and drug responses.

2. Disease research: Microarrays are utilized to identify genes associated with diseases such as cancer, neurodegenerative disorders, cardiovascular diseases, and autoimmune conditions. By comparing gene expression profiles between healthy and diseased tissues, researchers can identify potential biomarkers for disease diagnosis, prognosis, and treatment.

3. Pharmacogenomics: Microarrays are used to study how individuals respond to drugs based on their genetic makeup. By analyzing gene expression patterns in response to drug treatment, researchers can identify genetic factors that influence drug efficacy, toxicity, and individual variability in drug response. This information can inform personalized medicine approaches and improve drug development strategies.

4. Toxicogenomics: Microarrays are employed to study the effects of environmental toxins, chemicals, and drugs on gene expression patterns. By assessing changes in gene expression profiles in response to toxic exposures, researchers can identify potential biomarkers of toxicity, elucidate underlying mechanisms of toxicity, and assess the safety of chemicals and pharmaceuticals.

5. Agricultural biotechnology: Microarrays are used in plant research to study gene expression patterns associated with traits such as crop yield, stress tolerance, disease resistance, and nutritional content. This information can inform crop breeding programs, enhance agricultural productivity, and develop genetically modified crops with desirable traits.

Overall, gene microarrays are versatile tools that are widely used in research and diagnostics across various fields, including molecular biology, genetics, medicine, agriculture, and environmental science.

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The development of gene microarrays involved contributions from multiple researchers and laboratories over several years. However, one of the pioneering figures in the development of microarray technology is Dr. Patrick O. Brown, a biochemist and geneticist. In the early 1990s, Dr. Brown, along with colleagues at Stanford University, played a significant role in developing the concept and initial prototypes of DNA microarrays for studying gene expression patterns. Their groundbreaking work laid the foundation for the widespread adoption and further refinement of microarray technology in biological and medical research. While Dr. Brown's contributions were pivotal, it's important to note that the development of microarrays involved collaborative efforts from many scientists in the field.

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Northern Blot Gel Electrophoresis first uses denaturing gel to separate RNA according to its size.

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This procedure depends upon the process of nucleic acid hybridization between a known nucleic acid probe and the sequence that works with a population of RNA's.

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This type of analysis can be used to find out if a gene can be amplified, rearranged in cancer cells compared to noncancerous cells.

This technique if useful with detecting large deletions for tumor genomes.

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This method involves transferring DNA to a nylon membrane and probing it for presence of specific DNA sequences.

Different blood and tissue samples are used to detect changes in the DNA chains for irregular patterns.

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