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Merck
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Key Documents

D8276

Sigma-Aldrich

DNA Polymerase I, Klenow Fragment from Escherichia coli

buffered aqueous glycerol solution

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About This Item

CAS Number:
MDL number:
UNSPSC 코드:
12352204
NACRES:
NA.53

Grade

for molecular biology

Quality Level

형태

buffered aqueous glycerol solution

분자량

103 kDa

농도

~3,000 units/mL

UniProt 수납 번호

외래 활성

Endonuclease, none detected

배송 상태

wet ice

저장 온도

−20°C

유전자 정보

Escherichia coli K12 ... polA(948356)

일반 설명

DNA polymerase I yields two fragments (small and large) upon protease digestion. The large fragment (Klenow fragment) loses the 5′ exonuclease activity that is present in the intact holoenzyme. However, it retains both the polymerase 5′→3′ activity and the 3′→5′ exonuclease activity of the native enzyme.

애플리케이션

Suitable for:
  • DNA sequencing by the Sanger dideoxy method
  • Synthesis of the complementary strand of cDNA
  • Filling in 5′-overhangs in double stranded DNA to form blunt ends
  • Mutagenesis of DNA with second strand synthesis using oligonucleotides
  • Labeling DNA by the random primer method

성분

DNA Polymerase I is supplied as a solution in 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 5 mM dithiothretol, and 50% glycerol (v/v) .

단위 정의

One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min. at 37 °C.

재구성

The enzyme solution may be diluted with 50 mM Tris-HCl, pH 7.5, 100 mM ammonium sulfate, 10 mM 2-mercaptoethanol, and 1 mg/ml bovine serum albumin.

분석 메모

The activity is assayed in a reaction mixture containing 50 mM potassium phosphate (pH 7.5), 3 mM MgCl2, 1 mM 2-mercaptoethanol, 32.5 μM 32P-dATP, 32.5 μM dTTP, 62.5 μg/ml poly(dA-dT) and 0.01-1 unit enzyme.

픽토그램

Health hazard

신호어

Danger

유해 및 위험 성명서

예방조치 성명서

Hazard Classifications

Resp. Sens. 1

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point (°F)

Not applicable

Flash Point (°C)

Not applicable

개인 보호 장비

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


시험 성적서(COA)

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문서 라이브러리 방문

이미 열람한 고객

Sambrook, J.F., et al.
Molecular Cloning: A Laboratory Manual, 5-5 (1989)
R B Wallace et al.
Science (New York, N.Y.), 209(4463), 1396-1400 (1980-09-19)
Many eukaryotic genes contain intevening sequences, segments of DNA that interrupt the continuity of the gene. They are removed from RNA transcripts of the gene by a process known as splicing. The intervening sequence in a yeast tyrosine transfer RNA
DNA polymerase versus DNA binding to the anticancer drug, cis-platin.
Bose, R.N., et al.
Inorgorganica Chimica Acta, 300, 937-937 (2000)
Second-strand cDNA synthesis: mRNA fragments as primers.
U Gubler
Methods in enzymology, 152, 330-335 (1987-01-01)
Yossi Weizmann et al.
Journal of the American Chemical Society, 126(4), 1073-1080 (2004-01-30)
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