The percentage of methylation is not determined, however the enzymes used to produce this product completely cleave non-methylated lambda DNA. The methylated product is purified following this digestion process.
D3779
Lambda Phage DNA, Methylated from Escherichia coli host strain W3110
solution
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| A ustedes/SKU | Disponibilidad | Precio |
|---|---|---|
1 unit | Disponible para enviar HOYdesdeLyon | 50,20 € |
5 units | Póngase en contacto con nuestro Servicio de Atención al Cliente para disponibilidad | 127,00 € |
10 units | Póngase en contacto con nuestro Servicio de Atención al Cliente para disponibilidad | 246,00 € |
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Form:
solution
Storage temp.:
−20°C
50,20 €
Disponible para enviar HOYDetalles
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Molecular Biology
Quality Segment
form
solution
mol wt
31.5 × 103 kDa, 48 kb
concentration
556 μg/mL
suitability
suitable for substrate for restriction endonucleases
shipped in
dry ice
storage temp.
−20°C
General description
Phage DNA is isolated from infected E. coli, passed through a series of enzymatic steps before final phenol-chloroform extraction. This methylated lambda DNA is not completely digested by Bcl I, Cla I, Mbo I, Mbo II, Taq I or Xba I whereas non-methylated lambda DNA(product number D3654) is only partially cleaved.
Application
Lambda Phage DNA, Methylated from Escherichia coli host strain W3110 has been used:
- in magnetic tweezers measurements
- in the preparation of 35S-labeled DNA
- in the preparation of coupled standards (DNA + RNA)
Lambda Phage DNA, Methylated from Escherichia coli host strain W3110 is suitable for use as a substrate for restriction enzymes. It was used to study the attractive forces between condensed DNA double helix by combining single-molecule magnetic tweezers and osmotic stress. 35S-labeled Lambda Phage DNA, Methylated from Escherichia coli host strain W3110 was also used to standardize a method of measuring dissolved DNA in seawater.
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 W3110 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.
Infecting E. coli strain W3110 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.
Biochem/physiol Actions
Unique restriction sites: Apa I,Nae I, Nar I, Nhe I, PaeR7 I, SnaB I, Xba I and Xho I. Methylated lambda is partially cleaved by Bcl I, Cla I, Mbo I, Mbo II, Taq I and Xba I.
Physical form
DNA is supplied in a solution of 10 mM Tris-HCl (pH 8.0), with 1mM EDTA.
Other Notes
One O.D. (A260) is approximately 50 μg of DNA
1 of 1
Este artículo | |||
|---|---|---|---|
| description solution | description lyophilized powder | description solution | description aqueous glycerol solution |
| grade Molecular Biology | grade Molecular Biology | grade Molecular Biology | grade - |
| Quality Level 200 | Quality Level 200 | Quality Level 300 | Quality Level 200 |
| form solution | form lyophilized powder | form solution | form aqueous glycerol solution |
| concentration 556 μg/mL | concentration - | concentration - | concentration ≥2 unit/μL |
| storage temp. −20°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. −70°C |
| shipped in dry ice | shipped in - | shipped in dry ice | shipped in dry ice |
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Clase de almacenamiento
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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Is the sample "Lambda Phage DNA, Methylated from Escherichia coli host strain W3110 (D3779)" is fully (100% of A and C cites) methylated? If it is 100% methylated, how do you ensure the methylation percentage? Do you over express methylates?
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