XBA I
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XBA I 속성
- 저장 조건
- -20°C
- 물리적 상태
- 액체
안전
XBA I C화학적 특성, 용도, 생산
용도
The restriction enzyme Xba I has been used for the digestion of genomic DNA.일반 설명
Xba I recognizes the sequence T↓*CTAGA and generates fragments with 5′-cohesive termini. The enzyme needs at least two nucleotides around the target sequence before it will cut the cleavage site.Compatible ends
Xba I ends are compatible with fragments generated by Avr I, Nhe I, and Spe I.
Isoschizomers
The enzyme is not known to have isoschizomers.
Methylation sensitivity
Xba I digestion of DNA is inhibited by the dam gene product of E. coli, which methylates the 6N position of adenine in the sequence GATC. The enzyme is also inhibited by 5-methylcytosine or 5-hydroxymethylcytosine at the site (*) indicated on the recognition sequence.
Activity in SuRE/Cut Buffer System
Buffer printed in bold face type is the buffer recommended for optimal activity:
A B L M H
100% 75-100% 75-100% 75-100% 100%
Relative activity in complete PCR mix
Relative activity in PCR mix (Taq DNA Polymerase buffer) is 60%. The PCR mix contained ?DNA, primers, 10 mM Tris-HCl (pH 8.3, +20°C), 50 mM KCl, 1.5 mM MgCl2, 200 ?M dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. Activity in reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 25%. When supplemented with GC-RICH Solution, activity increases to 100%.
Incubation temperature
+37°C
Number of cleavage sites on different DNAs
λ Ad2 SV40 ?X174 M13mp7 M13mp18 pBR322 pBR328 pUC18
1 5 0 0 0 1 0 0 1
PFGE tested
Xba I has been tested in Pulsed-Field Gel Electrophoresis (on bacterial chromosomes). For cleavage of genomic DNA (E. coli C 600) embedded in agarose for PFGE analysis, we recommend 10 U enzyme per ?g DNA and approximately 4 hour incubation time.
Ligation and recutting assay
Xba I fragments obtained by complete digestion of 1 ?g pUC18 DNA are ligated with 1 U T4 DNA Ligase in a volume of 10 ?l by incubation for 16 hours at +4°C in 66 mM Tris-HCl, 5 mM MgCl2, 5 mM Dithiothreitol, 1 mM ATP, pH 7.5 (at +20°C) resulting in >90% recovery of pUC18 DNA.
Subsequent re-cutting with Xba I yields >95% of the typical pattern of pUC18 × Xba I fragments.
XBA I 준비 용품 및 원자재
원자재
준비 용품
XBA I 공급 업체
글로벌( 6)공급 업체
공급자 | 전화 | 이메일 | 국가 | 제품 수 | 이점 |
---|---|---|---|---|---|
FERMENTAS Inc. | -- |
United States | 335 | 82 | |
Amersham Biosciences, Inc. | -- |
United States | 1270 | 85 | |
Nacalai Tesque, Inc. | -- |
info-tech@nacalai.co.jp | Japan | 6048 | 75 |
Roche Applied Science | -- |
biochemts.us@roche.com | United States | 696 | 85 |
Qbiogene | -- |
orders@qbiogene.com | United States | 925 | 69 |
MP Biomedicals, Inc. | -- |
United States | 6566 | 81 |