Company Name: |
Qbiogene
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Tel: |
800 424 6101 |
Email: |
orders@qbiogene.com |
Products Intro: |
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Company Name: |
Nacalai Tesque, Inc.
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Tel: |
81 75 251 1723 |
Email: |
info-tech@nacalai.co.jp |
Products Intro: |
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Product Name: | XBA I | Synonyms: | XBA I | CAS: | | MF: | C97H123N38O58P9 | MW: | 3028 | EINECS: | | Product Categories: | | Mol File: | Mol File | |
| XBA I Chemical Properties |
storage temp. | -20°C | form | solution |
| XBA I Usage And Synthesis |
Uses | The restriction enzyme Xba I has been used for the digestion of genomic DNA. | General Description | 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 Preparation Products And Raw materials |
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