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Key Documents

D3654

Sigma-Aldrich

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

solution

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About This Item

CAS Number:
MDL number:
UNSPSC 코드:
41106310
eCl@ss:
32160414
NACRES:
NA.52

Grade

for molecular biology

Quality Level

형태

solution

분자량

31.5 × 103 kDa
48 kb

적합성

suitable for substrate for restriction endonucleases

배송 상태

dry ice

저장 온도

−20°C

유사한 제품을 찾으십니까? 방문 제품 비교 안내

특이성

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

애플리케이션

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.

물리적 형태

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.

기질

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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point (°F)

Not applicable

Flash Point (°C)

Not applicable

개인 보호 장비

Eyeshields, Gloves, type ABEK (EN14387) respirator filter


시험 성적서(COA)

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문서 라이브러리 방문

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Reclassification of `Pseudomonas fluorescens subsp. cellulosa?NCIMB 10462 (Ueda et al. 1952) as Cellvibrio japonicus sp. nov. and revival of Cellvibrio vulgaris sp. nov., nom. rev. and Cellvibrio fulvus sp. nov., nom. rev.
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International Journal of Systematic and Evolutionary Microbiology, 53(2), 393-400 (2003)
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