D8276
DNAポリメラーゼI, クレノウフラグメント 大腸菌由来
buffered aqueous glycerol solution
grade
Molecular Biology
form
buffered aqueous glycerol solution
mol wt
103 kDa
concentration
~3,000 units/mL
UniProt accession no.
foreign activity
Endonuclease, none detected
shipped in
wet ice
storage temp.
−20°C
Quality Level
Gene Information
Escherichia coli K12 ... polA(948356)
関連するカテゴリー
General description
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.
Application
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
Preparation Note
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.
Analysis Note
活性アッセイ:50mMリン酸カリウム, pH 7.5, 3mM MgCl2, 1mM 2-メルカプトエタノ-ル, 32.5μM32P-dATP, 32.5μM dTTP, 62.5μg/mLポリ(dA-dT), 0.01-1unit酵素。
Other Notes
1 unitは、37°C、30分間に、10 nmolのデオキシリボヌクレオシド三リン酸を酸性不溶性物質に変換する酵素量です。
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) .
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
保管分類
10 - Combustible liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
適用法令
試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。
D8276-BULK: + D8276-250UN: + D8276-50UN: + D8276-100UN: + D8276-VAR:
jan
H Klenow et al.
Proceedings of the National Academy of Sciences of the United States of America, 65(1), 168-175 (1970-01-01)
Purification of DNA polymerase from E. coli B has in two cases each time led to the isolation of two separate polymerase activities, enzyme A and enzyme B. Enzyme A was in contrast to enzyme B almost completely devoid of
Second-strand cDNA synthesis: mRNA fragments as primers.
U Gubler
Methods in enzymology, 152, 330-335 (1987-01-01)
L M Houdebine
Nucleic acids research, 3(3), 615-630 (1976-03-01)
E.Coli DNA polymerase I (Klenow subfragment) was used for the synthesis of complementary DNA with the mRNAs for rabbit milk proteins as templates. The cDNA formed, contained 200 nucleotides and represented about 20% of the mRNA template. The cDNA was
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)
関連コンテンツ
Instructions
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