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Cat No. Digestion site Quantity Isoschizomer
RD1001 5’...A G ↓C T...3’
3’...T C ↑ G A...5’ 60µl (1RDU/µl) AluBI
Source: Incubation Inactive Active site on λ DNA
Arthrobacter luteus 30 min at 37oC 20 min at 65oC 143
Supplied with: 300µl of 10X RapidDigest Universal Buffer,120 µl 10X RD Blue Buffer
Store at -20oC, avoid frequent thawing and freezing.
For in vitro use only
All RapidDigest Restriction Enzymes is completely active in Universal RD Buffer and digest DNA in 15-30 minutes or less.
RD restriction enzyme also eliminates need for sequential digestion during double digest methods.
1-Add below materials to 0.5ml tube:
Plasmid/ Lambda DNA PCR product Genomic DNA
Water DNase free 15ul 17ul 30ul
10X RapidDigest Buffer 2ul 2ul 5ul
DNA 2ul (up to 1ug) 10ul (˷0.2ug) 10ul (5ug)
RapidDigest Enzyme 1ul(1 RDU) 1ul(1 RDU) 5ul(5 RDU)
Total Volume 20ul 30ul 50ul
2- Mix gently and spin down.
3- Incubate at 37oC for 15-30 minutes1.
4- Inactive the enzyme by heating for 20 min at 65oC2.
1. Time of incubation may need to optimization but it could be achieved between 5 to 30 minutes. For digestion of difficult to cleave-DNA, incubation time may extend to one hour.
2. There are some alternative ways to stop the reaction:
- Addition of EDTA pH 8.0 <0.5M> final 20mM.
- Spin column DNA purification.
- Agarose gel extraction.
- Phenol- Chloroform extraction.
- Ethanol precipitation.
Ligation and recutting:
After 5-fold over digestion with, >95% of the DNA fragments can be ligated and recut with this enzyme.
No Inhibition: dam, CpG, dcm
One RapidDigest Unit (1 RDU) is the amount of enzyme required to completely digest 1µg of Lambda DNA in 15-30 minute in 1X RapidDigest buffer.
All preparations are assayed for contaminating endonuclease, 3'-exonuclease, 5'-exonuclease/
5'-phosphatase, as well as nonspecific single and double stranded DNase activities.
For double digestion simply add 1ul of each enzyme and scale up the reaction. For different temprature reaction start of the
enzymes that requires lower temprature.
λ DNA used as substrate for unit definition and quality control tests.