Genome DNA Analysis Grade Restriction Enzyme
Table of genome DNA for QC/Table showing amounts of enzymes required for complete digestion
When cutting various genome DNAs with restriction enzymes for the analysis of the genome DNAs, there is a wide variation of suitable restriction enzymes depending on the genome DNA species.
Takara selects suitable restriction enzymes for various genome DNA analysis, cuts appropriate bacterial genome DNA (agarose embedded; 0.5 µg DNA/50 µL gel) with each lot of the enzymes, and performs pulsed-field electrophoresis to confirm DNA bands pattern.
Amounts of enzymes required for complete digestion of bacterial genome DNA for QC was measured. To measure minimum amounts of enzyme required for complete digestion, 5, 10, 20 and 50 units of each enzyme was added and incubated for 5 and 20 hours.
Table of genome DNA for QC | Table showing amounts of enzymes required for complete digestion | |||||
---|---|---|---|---|---|---|
Restriction enzyme | Recognition sequences* | Genome DNA for QC | Reaction buffer** | Reaction temperature (°C) | Amounts of enzymes required for complete digestion | |
Incubation for 5 hrs |
Incubation for 20 hrs. |
|||||
Aat II | GACGTC | Staphylococcus aureus | T+BSA | 37 | 50 | 5 |
Aor 51H I | AGCGCT | Staphylococcus aureus | M | 37 | 50 | 20 |
Bgl I | GCCNNNNNGGC | Staphylococcus aureus | Basal | 37 | 10 | 5 |
Bln I | CCTAGG | Escherichia coli | K | 37 | 5 | 5 |
BspT104 I | TTCGAA | Arthrobacter luteus | L | 37 | 40 | 5 |
BssH II | GCGCGC | Staphylococcus aureus | M | 50 | 5 | 5 |
Bst1107 I | GTATAC | Arthrobacter luteus | K | 37 | 5 | 5 |
Cla I | ATCGAT | Arthrobacter luteus | M | 30 | 50 | 20 |
Cpo I | CGGWCCG | Staphylococcus aureus | K | 30 | 5 | 5 |
Dra I | TTTAAA | Pseudomonas aeruginosa | M | 37 | 100*** | 100*** |
Eco52 I | CGGCCG | Staphylococcus aureus | Basal | 37 | 10 | 5 |
Mlu I | ACGCGT | Staphylococcus aureus | H | 37 | 5 | 5 |
Mun I | CAATTG | Arthrobacter luteus | M+BSA | 37 | 5 | 5 |
Nae I | GCCGGC | Staphylococcus aureus | L | 37 | 50 | 50 |
Nhe I | GCTAGC | Arthrobacter luteus | M | 37 | 50 | 20 |
Not I | GCGGCCGC | Escherichia coli | H+BSA+Tri. | 37 | 5 | 5 |
Nru I | TCGCGA | Staphylococcus aureus | Basal | 37 | 50 | 5 |
Nsb I | TGCGCA | Arthrobacter luteus | T+BSA | 37 | 5 | 5 |
PshB I | ATTAAT | Pseudomonas aeruginosa | Basal | 37 | 5 | 5 |
Psp1406 I | AACGTT | Arthrobacter luteus | T+BSA | 37 | 5 | 5 |
Pvu I | CGATCG | Flavobacterium okeanokoites | K+BSA | 37 | 20 | 5 |
Sac II | CCGCGG | Staphylococcus aureus | T+BSA | 37 | 10 | 5 |
Sal I | GTCGAC | Staphylococcus aureus | H | 37 | 150 | 5 |
Sfi I | GGCCNNNNNGGCC | Escherichia coli | M | 50 | 100 | 50 |
Sma I | CCCGGG | Staphylococcus aureus | T+BSA | 30 | 5 | 5 |
Smi I | ATTTAAAT | Escherichichia coli | H | 30 | 50 | 5 |
SnaB I | TACGTA | Arthrobacter luteus | Basal | 37 | 10 | 10 |
Spe I | ACTAGT | Escherichia coli | M | 37 | 50 | 20 |
Sse8387 I | CCTGCAGG | Staphylococcus aureus | M+BSA | 37 | 5 | 5 |
Ssp I | AATATT | Arthrobacter luteus | Basal | 37 | 5 | 5 |
Xba I | TCTAGA | Escherichia coli | M+BSA | 37 | 50 | 5 |
Xho I | CTCGAG | Escherichia coli | H | 37 | 20 | 5 |
* W: A or T, N: A or C or G or T
** 0.5 µg DNA/50 µL gel+100 µL reaction mixture
*** Reaction of Dra I is extremely slow in agarose gel. Thus optimal amounts were added and left at 4°C for 16 hours. The samples were then transferred to 37°C and incubated for 5 or 20 hours.
References
- McClelland, M. (1987) Gene Amplification and Analysis 5 (Chirikijan, J. G. ed.), 257-282.2.
- McClelland, M., Patel, Y. and Nelson, M. (1987) Nucleic Acids Res., 15, 5985-6005.
- Daniels, D. L. (1990) Nucleic Acids Res., 18, 2649-2651.