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Pro. NATO. Acid. Sci. USA Vol. 88, pp. 1143-1147, February 1991 Biochemistry Single nucleotide primer extension to detect genetic diseases: Experimental application to hemophilia B (factor IX) and

How to fill out single nucleotide primer extension

How to fill out single nucleotide primer extension:

1. Begin by preparing a reaction mixture containing the DNA sample to be analyzed, a primer specific to the target sequence of interest, a labeled nucleotide (e.g., fluorescently labeled), DNA polymerase, and the necessary buffer components.
2. Denature the DNA sample and primer mixture by heating the reaction to a high temperature (typically around 95°C) for a short period (e.g., 5 minutes) and then quickly cooling it on ice. This step separates the DNA strands and allows the primer to anneal to the target sequence.
3. Add the labeled nucleotide to the reaction mixture, along with the DNA polymerase enzyme. The DNA polymerase will catalyze the addition of the labeled nucleotide to the primer, extending it by one nucleotide.
4. Incubate the reaction mixture at a specific temperature (typically around 72°C) for a fixed amount of time (e.g., 15 minutes) to allow the primer extension reaction to occur.
5. Stop the reaction by either adding a termination solution or by heating the reaction mixture to a high temperature to denature the DNA polymerase and stop any further incorporation of labeled nucleotides.
6. Analyze the products of the primer extension reaction using a method appropriate for the specific labeled nucleotide used (e.g., gel electrophoresis, capillary electrophoresis, or mass spectrometry). This will reveal the identity and relative quantity of the extended primers.

Who needs single nucleotide primer extension:

1. Researchers studying genetic variations or mutations: Single nucleotide primer extension is a valuable technique for analyzing specific genetic variants or mutations within a DNA sequence. It allows researchers to detect and quantify single nucleotide changes with high specificity and sensitivity.
2. Clinical laboratories performing genetic testing: Single nucleotide primer extension can be used in genetic diagnostic tests to identify disease-causing mutations or polymorphisms. This information can be crucial for diagnosing genetic disorders, guiding treatment decisions, and providing accurate genetic counseling.
3. Forensic laboratories: Single nucleotide primer extension can be used in forensic DNA analysis to identify specific genetic markers and variations. This technique is particularly useful in DNA profiling and individual identification.

Overall, single nucleotide primer extension is a versatile and powerful tool in molecular biology and genetics, enabling precise analysis of specific nucleotide sequences and genetic variations.

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What is single nucleotide primer extension?

Single nucleotide primer extension is a technique used in molecular biology to determine the exact composition of a DNA sequence at a specific location by extending a primer using fluorescently labeled nucleotides.

Who is required to file single nucleotide primer extension?

It is not specified who is required to file single nucleotide primer extension as it is a laboratory technique rather than a filing requirement.

How to fill out single nucleotide primer extension?

Single nucleotide primer extension is a laboratory technique and does not require any specific form or document to be filled out.

What is the purpose of single nucleotide primer extension?

The purpose of single nucleotide primer extension is to identify and analyze specific DNA sequences by determining the base composition at a particular location.

What information must be reported on single nucleotide primer extension?

There is no specific information that needs to be reported on single nucleotide primer extension as it is a laboratory technique rather than a reporting requirement.