D3654

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

solution

• CAS番号: 91080-14-7
• MDL番号: MFCD00146648
• UNSPSCコード: 41106310
• eCl@ss: 32160414
• NACRES: NA.52

特性

• グレード: for molecular biology
• 品質水準: 200
• フォーム: solution
• 分子量: 31.5 × 10³ kDa; 48 kb
• 適合性: suitable for substrate for restriction endonucleases
• 輸送温度: dry ice
• 保管温度: −20°C

詳細

特異性

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

アプリケーション

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.

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

• to calibrate spectrophotometer (LKB) and HPLC (high performance liquid chromatography) method for determination of bacterial genomic DNA G+C content
• to amplify the λ; exonuclease gene by polymerase chain reaction (PCR)
• in DNase activities

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.

物理的形状

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.

基質

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.

安全性情報

• 保管分類コード: 10 - Combustible liquids
• WGK: WGK 2
• 引火点(°F): Not applicable
• 引火点(℃): Not applicable
• 個人用保護具 (PPE): Eyeshields, Gloves, type ABEK (EN14387) respirator filter