• USA Home
  • D3654 - Lambda Phage DNA, Non-methylated from Escherichia coli host strain GM119 (rm-,dam-,dcm-)

EMAIL THIS PAGE TO A FRIEND
D3654 Sigma

Lambda Phage DNA, Non-methylated from Escherichia coli host strain GM119 (rm-,dam-,dcm-)

solution

Purchase

Properties

Related Categories Bacteriophage DNA, DNA, Molecular Biology, Molecular Biology Reagents
grade   for molecular biology
form   solution
mol wt   mol wt 31.5 × 103 kDa
  mol wt 48 kb
suitability   suitable for substrate for restriction endonucleases
shipped in   dry ice
storage temp.   −20°C

Description

Application

The lambda phage has an icosahedral head and a long tail terminating in a single fiber. At both ends of the 5′ termini are complementary 12-nucleotide single strand sequences that contribute to the cohesive ends (cos region) of the DNA. The tail of the phage latches on the host outer membrane receptor and injects phage DNA into the cell. The phage converts the E. coli to a lysogenic state in which the phage functions are repressed and the phage genome may remain dormant (prophage) for a long time. This property is seen in bacteriophages that carry CII and CIII genes that are responsible for CI expression. Bacteriophages with CI mutation in the CI gene are able to maintain a lysogenic state at defined temperatures.
Infecting E. coli strain GM 119 with lambda C1857 strain creates E. coli lysogen cultures. The phage is released from E. coli cell pellets by lysing with a high salt buffer, pH 8.0. The crude mixture is passed through a series of enzymatic steps, multiple cesium gradients, and phage DNA is dialyzed against 1 mM Tris-HCl, pH 8.0, and 1 mM magnesium chloride. The DNA is finally extracted by phenol-chloroform solution.

Lambda Phage DNA, Non-methylated from Escherichia coli host strain GM119 is suitable for use as a substrate for restriction enzymes. It was used as substrate for Prevotella ruminicola DNase activity studies. It was also used as a GC content standard in genome sequencing of 16S rDNA sequences of Cellvibrio japonicas.

Specificity

Unique Restriction Sites (Methylated DNA) are: Apa I,Nae I, Nar I, Nhe I, PaeR7 I, SnaB I, Xba I and Xho I.

Substrates

Non-methylated lambda DNA is completely digested by Bcl I, Cla I, Mbo I, Mbo II, Taq I or Xba I whereas methylated lambda DNA is only partially cleaved.

Physical form

Phage DNA is isolated from infected E. coli, passed through a series of enzymatic steps before final phenol-chloroform extraction. This non-methylated lambda DNA is completely digested by Bcl I, Cla I, Mbo I, Mbo II, Taq I or Xba I whereas methylated lambda DNA(product number D3779) is only partially cleaved. The unique restriction sites are Apa I,Nae I, Nar I, Nhe I, PaeR7 I, SnaB I, Xba I and Xho I.

Price and Availability

Protocols & Articles
Peer-Reviewed Papers
15

References

Set your institution to view full text papers.
Related Products

related product

Product #

Description

Add to Cart

D9768 Lambda Phage DNA, Methylated from Escherichia coli host strain W3110, lyophilized powder
D3779 Lambda Phage DNA, Methylated from Escherichia coli host strain W3110, solution

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?