How is M13 DNA isolated from infected cells?

Bacteriophage M13 single-stranded DNA is prepared from virus particles secreted by infected bacteria into the surrounding medium. Subsequent extraction with phenol releases the single-stranded DNA, which is then collected by precipitation with ethanol.

How do you extract DNA from bacteria at home?

DNA EXTRACTION FROM BACTERIA

1. Write your name or initials on a test tube containing detergent. Pour the E.
2. (Optional) Remove the tube from the hot water bath.
3. Remove the tube from the hot water bath.
4. Let the solution sit for 2-3 minutes without disturbing it.

How do you extract bacteriophages?

The isolation of bacteriophages for phage therapy is often presented as a fairly straightforward exercise of mixing a phage-containing sample with host bacteria, followed by a simple removal of bacterial debris by filtration and/or centrifugation the next day [1,2,3].

How is bacteriophage DNA purified?

The phage DNA is preferentially purified from other cellular components such as proteins without the use of phenol, chloroform or cesium chloride. The starting material for this procedure is clarified phage supernatant that has been separated from bacterial debris in liquid cultures.

How does bacteriophage M13 replicate?

Additional single-stranded copies of the M13 genome are made through rolling circle replication using the RF DNA as a template. While concentrations of phage proteins are low, the single-stranded copies are converted into additional RF DNA.

Why is M13 a clone vector?

M13 cloning vectors have been developed for the selection of DNA sequences capable of directing initiation of DNA synthesis on single-stranded templates. These vectors are derived from viable M13 mutants containing large deletions in the region of the complementary strand origin.

What does salt solution do to extract DNA?

The salt neutralizes the negative charges on the DNA and thus enables the DNA strands to stick together. It also causes proteins and carbohydrates to precipitate.

How do you extract DNA phage?

The best methods to obtain the maximum yield of DNA were precipitation of phage with ultracentrifugation or acetone precipitation followed by extraction by commercial kit, potassium iodide or sodium iodide extraction.

Where are bacteriophages found?

Bacteriophages are viruses that infect bacteria. Also known as phages (coming from the root word ‘phagein’ meaning “to eat”), these viruses can be found everywhere bacteria exist including, in the soil, deep within the earth’s crust, inside plants and animals, and even in the oceans.

How can bacteriophage DNA be spread?

Bacteriophages work in different ways; some enter their bacterial host and incorporate their genome into the bacterial DNA, happy to settle down and replicate with the host. Others multiply inside the bacteria to create new phage genomes, which then burst out of the host and spread.

What is the size of the DNA molecule which can be packed by the bacteriophage M13 protein coat?

6407 nucleotides
Ff phages are composed of circular single-stranded DNA (ssDNA) which is 6407 nucleotides long encapsidated in approximately 2700 copies of the major coat protein p8, and capped with about 5 copies each of four different minor coat proteins (p3 and p6 at one end and p7 and p9 at the other end).