DNA Concentration Measurement Methods

<p>Protein or phenol contamination (C)</p> Signup and view all the answers

<p>5 to 50 µL (B)</p> Signup and view all the answers

<p>To separate nuclear and mitochondrial DNA (C)</p> Signup and view all the answers

<p>Warming up the UV-lamp (C)</p> Signup and view all the answers

<p>Uniform mixing of the sample (B)</p> Signup and view all the answers

<p>Chelate Mg 2+ to inactivate nucleases (D)</p> Signup and view all the answers

<p>Alkaline buffer (A)</p> Signup and view all the answers

<p>Separates contaminants into organic phase (A)</p> Signup and view all the answers

<p>Only negatively charged molecules move towards the positive electrode. (D)</p> Signup and view all the answers

<p>Isopropanol (B)</p> Signup and view all the answers

<p>For staining DNA to visualize it (B)</p> Signup and view all the answers

<p>0.04 μg/μL corresponds to 40 μg/mL (B)</p> Signup and view all the answers

<p>Charge, size, and shape (D)</p> Signup and view all the answers

<p>It provides density so the samples can sink into the wells. (B)</p> Signup and view all the answers

<p>Ethidium bromide (A)</p> Signup and view all the answers

<p>It conducts electricity and controls pH. (D)</p> Signup and view all the answers

<p>They move through the pores of the gel more easily. (C)</p> Signup and view all the answers

<p>0.8 g (A)</p> Signup and view all the answers

<p>Bromophenol blue (B)</p> Signup and view all the answers

<p>Negative charge (D)</p> Signup and view all the answers

<p>1500 mL (C)</p> Signup and view all the answers

<p>To visualize nucleic acids under UV light (A)</p> Signup and view all the answers

<p>55°C (A)</p> Signup and view all the answers

<p>To halt enzymatic reactions (D)</p> Signup and view all the answers

<p>To prevent damage from ultraviolet light (B)</p> Signup and view all the answers

<p>Sodium bicarbonate (C)</p> Signup and view all the answers

<p>STBS has a higher pH than TAE (C)</p> Signup and view all the answers

<p>0.5 µg/mL (B)</p> Signup and view all the answers

<p>Contains Tris-Cl and NaEDTA, pH 8 (A)</p> Signup and view all the answers

<p>They migrate slower (B)</p> Signup and view all the answers

<p>50 mM (C)</p> Signup and view all the answers

<p>Add 1 µL of gel loading dye for every 5 µL of DNA solution (A)</p> Signup and view all the answers

<p>20 minutes (D)</p> Signup and view all the answers

<p>The gel should be stained in 0.5 µg/mL ethidium bromide (B)</p> Signup and view all the answers

<p>To enhance the resolution of different sizes of DNA fragments (D)</p> Signup and view all the answers

<p>It allows larger fragments to migrate faster than smaller ones (D)</p> Signup and view all the answers

<p>DNA migration will effectively cease (B)</p> Signup and view all the answers

<p>Tris-borate-EDTA (TBE) (B)</p> Signup and view all the answers

<p>It may melt the gel due to excessive heat generation (D)</p> Signup and view all the answers

<p>To aid in the visibility and density of DNA samples for loading (C)</p> Signup and view all the answers

<p>1% final (B)</p> Signup and view all the answers

<p>They influence DNA migration rates (A)</p> Signup and view all the answers

Determining the amount of DNA in a sample, crucial for subsequent analyses.

A method for measuring DNA concentration by quantifying UV light absorption by DNA in solution.

50 µg/mL of dsDNA absorbs 1.0 at 260 nm.

40 µg/mL of ssDNA or RNA absorbs 1.0 at 260 nm.

Indicates DNA purity. Pure DNA has ratio ~1.8. Lower ratios suggest contaminants.

Used for assessing DNA quantity and quality (size, degradation), and RNA presence.

Volume of DNA sample needed for spectrophotometric analysis (5-50 µL).

Genomic DNA ~100 µg/mL, plasmid DNA ~1 mg/mL (Approximate values).

A polysaccharide used to create a gel matrix in electrophoresis for molecule separation

The movement of charged molecules in a gel matrix under the influence of an electric field.

Dye added to samples to improve visualization and density during electrophoresis using glycerol or sucrose

A method including lysing cells, removing polysaccharides, proteins and separating DNA using specific buffers for DNA precipitation.

Measuring the amount of RNA in a solution, 40 µg/mL = 1.0 absorbance unit at 260 nm.

A buffer used to solubilize cells and complex with cellular nucleic acids, helping separate nucleic acid.

A solution having an almost constant pH, used in an electrophoresis apparatus to carry current and maintain the pH of separating solutions. Examples include TAE, TBE, TPE, or alkaline buffers.

A solution that conducts electricity and controls pH, essential for DNA movement in electrophoresis.

A solution mixed with DNA samples to make them denser and help them sink into the gel wells.

A DNA-binding dye used to stain DNA fragments, making them visible under UV light.

Process of making an agarose gel for separating genomic DNA, using 0.8g Agarose.

Process of making an agarose gel for separating PCR product, using 1.5-2 g Agarose.

Indentation in the agarose gel where DNA samples are loaded.

Movement of negatively charged DNA fragments towards the positive electrode in an electric field.

An ultraviolet lightbox used to visualize DNA stained with ethidium bromide.

Dissolving agarose powder in a buffer, allowing solidification in a tray to form a gel matrix for electrophoresis.

Linear DNA fragments in an agarose gel migrate inversely proportional to the logarithm of their molecular weight or base pairs.

Adding ethidium bromide to the gel or electrophoresis buffer after electrophoresis to visualize the DNA bands.

A buffer commonly used for extracting DNA, containing Tris-Cl, NaCl, and EDTA.

A solution containing sucrose, Triton X-100, EDTA, and Tris-Cl, used for lysis and cell disruption.

A solution used to suspend and store bacteria, containing NaCl, MgSO4, Tris-Cl, gelatin, and water.

A buffer used for ligating DNA fragments, containing Tris-Cl, MgCl2, and DTT.

A buffer used for cloning and transformation reactions, containing Tris-acetate, potassium acetate, magnesium acetate, DTT, and BSA.

The percentage of agarose in a gel determines which size DNA fragments are best separated.

Increasing voltage makes larger DNA fragments migrate faster, impacting separation.

Buffers provide ions for conductivity and maintain a stable pH during electrophoresis.

Using a buffer is critical for DNA migration in gels. Without it, DNA won't move.

Using a concentrated buffer can generate excessive heat, potentially melting the gel.

A solution containing tracking dyes and sucrose, added to DNA samples before loading onto a gel.