EcoR I info

Mi Fermentas #

CERTIFICATE OF ANALYSIS

EcoBl

#ER0272    5x5000u

5’...G^A A T T C...3'

3-...C T T A At G...5-

Lot    2911

Ouality guaranteed: 02.2005 Concentration:    10 unfts/pl

Source:    E.coli that carries the cloned

ecoRIRgene from Escherichia coli RY13

Storę at -20°C

B ® ą B S ® 0 ® § lllllllllll liliilllllllllll SSS.

0002722911020528

wwwifermentas.com

UTKUANIA tel.:[phone]. fax:[phone], infoOfermentas.lt germany u^>l.:+49 62 27/55853, fax:[phone], fermentasOt-onHne.de usa tel.: [phone], fax: [phone], lnfoOfermentas.com canada tel.: [phone], fax: [phone], bifoOfermentas.com

RECOMMENDED CONDITIONS Buffer EcoRl*

50mM Tris-HCI (pH7.5), 10mM MgCI_?, 100mM NaCI, 0.02% Triton X-100,0.1 mg/ml BSA.

Incubate at 37°C.

Unit Definitio n

One unit is defined as the amount of enzyme required to digest 1 pg of lambda DNA in 1 hour at 37°C in 50pl of assay buffer.

Dilution

For short-term storage (3-4 weeks) - dilute with Diluent Buffer (#B19): 10mM Tris-HCI (pH7.4 at 25°C),

10OmM KCI, 1 mM EDTA, 1 mM DTT, 0.2mg/ml BSA and 50% glyceroi.

For longer periods - the Storage Buffer should be used. Double Digests

Y7TANqo™ Buffer is provided to simplify buffer selection for double digests. 98% of Fermentas Restriction Endonucleases (REs) work well in a 1x or 2x concentration of the YTTanoo™ Buffer. Please refer to the Enzyme Activity chart (see enzyme properties) to choose the best buffer for the two REs in your digest. Storage Buffer

10mM potassium phosphate (pH7.4 at 25°C),

300mM NaCI, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA, 0.15% Triton X-100 and 50% glyceroi.

QUALITY CONTROL ASSAY DATA

Overdigestion Assay

No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/pg lambda DNA x 16 hours) with feoRI (see Star ActMty).

Ligation/Recutting Assay After 50-fold overdigestion (3u/pg DNA x 17 hours) with fcoRI, morę than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.02pM. Morę than 95% of these can be recut.

Labeled Oligonucleotide (LO) Assay No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours.

Blue/White Cloning Assay pUC57 was digested at a unique site with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.

Ouality authorized by:    Jurgita Zilinskiene

PRODUCT USEUMITAT10N.

Tlils product Is developed. daslgned and sold exdusively lor research purposes and In vilm use only. The producl was not lesled tor use In diagnostics or for drug davelopment, nor Is II sultabłe lor adminlsliaiion to humans ot anlmals.

Updated November 13.2002

ENZYME PROPERTIES

Enzyme Activity in Fermentas Buffers, %

fcoRI* B* 6* O* B* YVr«.qo’“

__IX 2X

100    0-20 NR 100    100* m 100

'Star actmty appears at 10-fold overdigestion (10 units x 1 houi).

NR: Buffer is not recommended, sińce star acfcity appears at 5-foid overdigest!on (5 units x 1 hou).

StarActivity

A large excess of enzyme (1 Ou/pg DNA x 16 hours), Iow salt concentration, high pH, or the replacement of Mg²* by Mn²* may result in star actMty.

Stability during Prolonged Incubation A minimum of 0.2 units of enzyme is required for complete digestion of 1pg of lambda DNA in 16 hours at 37°C.

Thermal lnactivation

Enzyme is inactivated by incubation at 65°C for 20 min. Digestion of Agarose-embedded DNA A minimum of 5 units of enzyme is required for complete digestion of 1pg of agarose-embedded lambda DNA in 16 hours.

Compatible Ends Xap\, Mun\, TasI

Number of Recognition Sites in DNA

).    <I»X174 pBR322 pUC57 pUC18/19 pTZI9R/U M13mp18/19