Rapid Digest BglII

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Rapid Digest BglII

Cat No.

Digestion site

Quantity

Isoschizomer

RD1021

5’...A ↓G A T C T ...3’

3’...T C T A G ↑A...5’

60µl(1RDU/µl)

-

source

Incubation time

Inactive

Active site on  λ DNA

Bacillus globigii 

30 minutes at 37oC

-

6

 

 

 

 

 

 

 

 

Supplied with:            300µl 10X RD Universal Buffer, 120µl 10X RD Blue Buffer

               

Store at -20oC, avoid frequent thawing and freezing.

For in vitro use only

Rapid Digest Restriction Enzyme is completely active in Universal Rapid Digest Buffer. All RD restriction enzymes are able to digest DNA in 15-30 minutes or less.

RD restriction enzyme also eliminates need for sequential digestion during double digest methods.

 

Recommended assay

1-Add below materials to 0.5ml tube:

 

 

Plasmid/ Lambda DNA

PCR product

Genomic DNA

Water DNase free

15ul

17ul

30ul

10XRapidDigest Buffer

2ul

2ul

5ul

DNA

2ul (up to 1ug)

10ul (˷0.2ug)

10ul (5ug)

Rapid Digest Enzyme

1ul

1ul

5ul

Total Volume

20ul

30ul

50ul

 

2- Mix gently and spin down.

3- Incubate at 37oC for 30 minutes1.

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.

 

Ligation and recutting:

After 50-fold over digestion with, >95% of the DNA fragments can be ligated and recut with this enzyme.

 

DNA Methylation:

No Inhibition: dcm, dam, CpG

 

       Unit Definition:

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. 

 

Quality Control:

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.