컨텐츠 바로가기 영역
주메뉴로 바로가기
본문으로 바로가기

Restriction Endonuclease

Home > PRODUCTS > Restriction Endonuclease

Apa I

No Product Name Cat.# Conc Size Price(₩) Add
1 Apa I R020S 8 units/µl 2,000 units ₩ 57,000 추가
2 Apa I R020M 8 units/µl 4,000 units ₩ 102,600 추가
3 Apa I R020L 8 units/µl 10,000 units ₩ 228,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
Total : 장바구니에 담기 바로 구매 하기
plus minus 삭제

Recognition & cleavage sequence


Source: Acetobacter pasteurianus sub pasteurianus

 

Reaction conditions

1X EzBuffer IV, 25

1X FastCut Buffer, 25

 

FastCut Buffer

Enzynomics restriction enzyme can cut substrate DNA in 5-15 min with FastCut Buffer

 

1X EzBuffer IV

20 mM Tris-acetate (pH 7.9 @ 25), 50 mM potassium acetate, 10 mM magnesium acetate, 100 μg/ml BSA

 

Unit definition

One unit is defined as the amount of enzyme required for complete digestion of 1-μg bacteriophage λ at 25 for 1 hr in 50-μl reaction mixtures.

 

Storage

10 mM Tris-HCl (pH 7.4 @ 25), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA, 50% glycerol.

 

Dilution buffer: EzDiluent A

10 mM Tris-HCl (pH 7.4 @ 25), 50 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA, 50% glycerol.

 

Heat Inactivation

Apa I can be inactivated at 65 for 20 min.

 

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Conditionally sensitive

CpG methylation: Conditionally sensitive

 

Prolonged incubation

A minimum amount of enzyme required to digest 1-μg substrate DNA for 16 hr; 0.13 U.

 

Relative activity in EzBuffers

EzBuffer I: 100%

EzBuffer II: 25%

EzBuffer III: 0%

EzBuffer IV: 100%

FastCut Buffer: 100%

 

Note

Incubation at 37 results in 100% activity. However, it is unstable at 37 with a half-life of only 30 min. Either dcm or CpG methylation overlapping its recognition sequence blocks cleavage of DNA. Salt over 50 mM is inhibitory.

● Material Safety Data Sheet

- Enzynomics_MSDS_R020_Apa I

 

● Quality control

Overdigestion assay

DNA digested for 16 hr in 50-μl reaction with 100 U of enzyme resulted in same the DNA band patterns as those obtained with 1 U of enzyme for 1 hr.

 

Endonuclease assay

Less than 5% of 1-μg of ΦX174 RFI is converted to RFII when the DNA is incubated with 50 U of enzyme at 37 for 4hr in 50-μl reaction. 

 

Ligation and recutting

More than 95% of DNA fragments (1 μg) digested with 50-fold excess enzyme can be ligated by T4 DNA ligase (400 U) at 16 for 16 hr. Of the ligation products, > 95% can be re-cut. 

 

Extreme pure (EP)

No detectable degradation of 32P-end labeled single-stranded and double-stranded (5’-, 3’-overhang and blunt end) oligonucleotides occurred during incubation with 100 U of enzyme for 4 hr.

 

List