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
Restriction enzyme	Recognition sequences*	Genome DNA for QC	Reaction buffer**	Reaction temperature (°C)	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.

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