Lambda Phage DNA, Methylated fromEscherichia colihost strain W3110

半岛bd体育手机客户端 说明

一般描述

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.

特异性

Unique restriction sites:ApaI,NaeI,NarI,NheI,PaeR7 I,SnaBI,XbaI andXhoI. Methylated lambda is partially cleaved byBclI,ClaI,MboI,MboII,TaqI andXbaI.

应用

Lambda Phage DNA, Methylated fromEscherichia colihost 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 fromEscherichia colihost strain W3110 was also used to standardize a method of measuring dissolved DNA in seawater.
Lambda Phage DNA, Methylated fromEscherichia colihost strain W3110 has been used:

• in magnetic tweezers measurements
• in the preparation of³⁵S-labeled DNA
• in the preparation of coupled standards (DNA + RNA)

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 ( cosregion) 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. colito 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. colistrain W3110 with lambda C1857 strain creates E. colilysogen cultures. The phage is released from E. colicell 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.

单位定义

One O.D. (A₂₆₀) is approximately 50 μg of DNA

外形

DNA is supplied in a solution of 10 mM Tris-HCl (pH 8.0), with 1mM EDTA.

基本信息

MDL编号	MFCD00146648
NACRES	NA.52

半岛bd体育手机客户端 性质

质量水平	200
等级	for molecular biology
形式	solution
分子量	31.5 × 103kDa 48 kb
浓度	556 μg/mL
适用性	suitable for substrate for restriction endonucleases
运输	dry ice
储存温度	?20℃

安全信息

储存分类代码	12 - Non Combustible Liquids
闪点(F)	Not applicable
闪点(C)	Not applicable