What is restriction digestion of lambda DNA?
Since the lambda genome is approximately 48,000 base pairs, each restriction enzyme will cut the DNA several times and generate restriction fragments of different sizes. In this activity, three separate samples of lambda DNA will be cut using three different restriction enzymes, and one sample will remain undigested.
Which lane shows a digested DNA sample?
Lane 4: Digested PCR product (or DNA Fragment).
How many bands are visible in the digested lambda DNA?
Lambda phage-DNA has 5 EcoRI restriction sites so after it should produces 6 bands but I can see only 4 out of 6 possible bands.
What is lambda repressor?
Introduction. cI is a transcription inhibitor of bacteriophage Lambda. Also known as Lambda Repressor, cI is responsible for maintaining the lysogenic life cycle of phage Lambda. This is achieved when two repressor dimers bind cooperatively to adjacent operator sites on the DNA.
How do you digest plasmid DNA?
- Select restriction enzymes to digest your plasmid.
- Determine an appropriate reaction buffer by reading the instructions for your enzyme.
- In a 1.5mL tube combine the following:
- Mix gently by pipetting.
- Incubate tube at appropriate temperature (usually 37 °C) for 1 hour.
How is the lambda DNA marker digested and purified?
Lambda DNA is digested to completion with the appropriate Thermo Scientific restriction enzyme (s) and purified and dissolved in storage buffer. DNA fragments contain various sticky ends depending on the restriction enzyme used for preparation of the marker. The marker can be labeled radioactively with T4 Polynucleotide Kinase.
Are the λ DNA/HindIII markers intended for quantitative analysis?
These markers are not intended for quantitative analysis. The λ DNA/HindIII Markers have eight ethanol-precipitated DNA fragments ranging in size from 125bp to 23,130bp. Storage Buffer: 10mM Tris-HCl (pH 7.8), 10mM NaCl, 1mM EDTA.
How many DNA fragments does HindIII digest of lambda DNA?
The HindIII digest of lambda DNA ( c I857 ind 1 Sam 7) yields 8 fragments suitable for use as molecular weight standards for agarose gel electrophoresis (1). The approximate mass of DNA in each of the bands is provided (assuming a 1.0 μg load) for approximating the mass of DNA in comparably intense samples of similar size.
How is DNA-HindIII digest visualized in lambda?
Lambda DNA-HindIII Digest visualized by ethidium bromide staining. 1.0% agarose gel. The full list of DNA fragments can be found in the chart below.