What is the purpose of uncut lambda DNA?
Lambda DNA (48,502 bp) may be used as a molecular weight size marker during nucleic acid gel analysis following digestion with a restriction enzyme (such as HindIII). Lambda DNA can also be used as a substrate in restriction enzyme activity assays.
What is Lambda HindIII DNA marker?
Thermo Scientific Lambda DNA/HindIII Marker is recommended for sizing of linear double-stranded large DNA fragments in agarose gels. Lambda DNA is digested to completion with the appropriate Thermo Scientific restriction enzyme(s) and purified and dissolved in storage buffer.
How many base pairs long is Lambda?
49130 base pairs
Phage lambda DNA is a double-stranded, linear molecule, 49130 base pairs in length.
What are all of the sizes of the pieces you expect if you cut lambda DNA with HindIII alone?
Cutting with Hin dIII will yield fragments of 1, 2, and 2.5 kilobases. Cutting with Eco RI will yield fragments of 1.5, 2, and 3.5 kilobases.
How many fragments will HindIII cut lambda DNA into?
The HindIII digest of lambda DNA yields at least 6 fragments suitable for use as molecular weight standards for gel electrophoresis.
How many ecor1 sites are there in lambda DNA?
5 recognition sites
The Lambda DNA used in this experiment is isolated as a linear molecule from the E. coli bacteriophage lambda. It contains approximately 49,000 base pairs and has 5 recognition sites for Eco RI, and 7 for Hind III.
Is lambda DNA double stranded?
The virion DNA is linear and double-stranded (48502 bp) with 12 bp single-stranded complementary 5′-ends. After the phage particle injects its chromosome into the cell, the chromosome circularizes by end joining.
How does gel electrophoresis analyze DNA?
Gel electrophoresis and DNA DNA is negatively charged, therefore, when an electric current is applied to the gel, DNA will migrate towards the positively charged electrode. Shorter strands of DNA move more quickly through the gel than longer strands resulting in the fragments being arranged in order of size.
How do you check DNA quality on gel?
To evaluate DNA purity, measure absorbance from 230nm to 320nm to detect other possible contaminants. The most common purity calculation is the ratio of the absorbance at 260nm divided by the reading at 280nm. Good-quality DNA will have an A260/A280 ratio of 1.7–2.0.
When is a concatemer not formed in a cell?
Interestingly, concatemers are not formed when E. coli RNA polymerase is not adequately inhibited. For example, during T7Δ2 infection of E. coli, no high-molecular weight T7 DNA is found in the cells [54,55].
How are concatemers formed on the T7 chromosome?
In vivo [51] and in vitro [52,53] studies suggest that concatemers are formed through the annealing of the 3′-ends of the terminally redundant tails of the T7 chromosome that are generated by DNA replication. Interestingly, concatemers are not formed when E. coli RNA polymerase is not adequately inhibited.
What does DNA gel look like under a microscope?
Smaller DNA fragments can move quickly through the pores, while larger fragments get caught and therefore travel slowly. Let’s look at how this all works. Under a powerful microscope, a gel will look porous, but to the naked eye, it looks like a smooth, opaque gelatin in the shape of a square with wells near one end of the surface.
How are concatemers used in the replication process?
A concatemer is a molecule made up of multiple copies of the same genome strung together in tandem. For viroids and many bacteriophages, this sort of structure is a standard part of the replication process. Viroid reproduction involves the formation of linear concatemers of single-stranded DNA,…