Details

Source

Chlorella virus IL-3ANote:Purified from a recombinant source (Patent No. US005472872A)

Reagents Supplied

10X CviJ I* Reaction BufferDMSO

Description

• CviJ I* is an unique restriction enzyme capable of digesting DNA at two or three base recognition sequence (1,2)
• CviJ I (Cat. No. 2125-01) normally cleaves the sequence 5...PuGCPy...3 between the G and C to leave blunt ends
• Under \"relaxed\" conditions (in the presence of Mg2+, ATP and enhancers), CviJ I* cleaves the sequences 5...GC...3 except 5...PyGCPu...3
• Capable of cleaving single-stranded DNA and double-stranded DNA into small 20-200 bp fragments
• Generates numerous sequence specific oligonucleotides from unknown DNA samples
• Exclusively from CHIMERx

Applications

• CviJ I and CviJ I* cleave DNA extremely frequently and thus can be used for a variety of novel molecular biology applications (2,3,4)
• CviJ I / CviJ I* partial digests can also be used in applications such as shot-gun cloning, generating quasi-random libraries (2) and epitope mapping or panning
• CviJ I digestion of anonymous DNA produces a large number of oligonucleotide sized polymers upon thermal denaturation, which can be exploited in applications such as:

Incubation Temperature

37°C

Heat Inactivation

65°C for 10 minutes

Assay Unit Substrate

pBR322

Reaction Buffer

10X CviJ I* Reaction Buffer:20 mM glycyl-glycine KOH (pH 8.5)10 mM magnesium acetate50 mM potassium acetate0.1 mM ATP0.1 mM dithiothreitolNote: 100%DMSO supplied separately

Storage Buffer

20 mM Tris-acetate (pH 8.0 at 4°C)0.5 mM EDTA0.1 mM dithiothreitol5 mM magnesium chloride50 mM potassium acetate50% (v/v) glycerol

Assay Conditions

20 mM glycylglycine-KOH (pH 8.5) 10 mM magnesium acetate0.1 mM dithiothreitol50 mM potassium acetate0.1 mM ATP 30% DMSO 1 µg pBR322Incubation for 3 hours at 37°C in a total reaction volume of 25 µl

Storage Conditions

Store at -80°CShipped on dry ice

Customer Note(s)

(1) CviJ I* restriction endonuclease is inhibited by glycerol concentrations in excess of 2.5%. Therefore the extension of the digestion time is recommended rather than using additional units of CviJ I*. Alternatively, DNA sample can be ethanol precipitated and re-digested.

(2) Due to extreme frequency of CviJ I*/ CviJ I recognition sites, sterical interference of closely located recognition sites is observed. It results in slower digestion of such sites. In consequence, the generated oligonucleotide fragments are rarely shorter than 15 bp that makes them ideal for anonymous primer applications.

(3) CviJ I* reaction buffer contains DMSO, which does not interfere with further enzymatic manipulations (ligations, labeling, etc). If the sample is intended for electrophoresis, ethanol precipitatation of the reaction mixture after completed digestion is strongly recommended in order to avoid diffused bands on agarose or polyacrylamide gels.

Downloads

Certificate of Analysis PDF-Current LotMSDS PDF-Current Lot

References

(1) Xia, Y., Burbank, D., Uher, L., Rabussay, D. and Van Etten, J. Nucleic Acids Res.15, 6075-6090(2) Fitzgerald,M.C., Skowron, P., Van Etten, J.L., Smith, L.M. and Mead, D.A. (1992) Nucleic Acids Res. 20,3753-3762(3) Skowron, P.M, Swaminathan, N., McMaster, K., George, D., Van Etten, J. andMead, D. Gene 157 (1995) 37-41(4) Mead, D., Swaminathan, N., Van Etten J. and Skowron, P.M.: Recombinant CviJI restriction endonuclease. (1995) Unites States Patent no US005472872A(5) Swaminathan, N., McMaster, K., Skowron, P. and Mead, D.A. Analytical Biochemistry 255 (1998) 133-141