Deoxyribonuclease I (DNase I) , GMP Grade

DNase I is an endonuclease that can digest single-stranded and double-stranded DNA to produce single deoxynucleotides or single-stranded ordouble-stranded oligodeoxynucleotides.It can hydrolyze the phosphodiester bond to produce monodeoxynucleotidesand oligodeoxynucleotides containing 5′-phosphate groups and 3′-OH groups.The average digestion product is the smallest polytetranucleotide.DNasel can catalyze many forms of DNA, such as single-stranded DNA,double-stranded DNA,and even chromatin(its cutting rate is affected by histones).The optimum pH range is 7-8.The activity of DNase I depends on Ca²+and can be activated by divalent metal ions,suchas Co2+,Mn²+,Zn²+,etc.5mM Ca²+can protect the enzyme from being hydrolyzed. In the presence of Mg ²+,the enzyme can recognize and cut any site on any strand of DNA randomly;and in the presence of Mn²+,it can recognize two strands of DNA at the same time and cut at almost the same site to form blunt ends,Or sticky ends with 1-2 nucleotides protruding.DNase lis widely used in the preparation of DNA-free RNA;removethe template DNA after in vitro transcription;prepare DNA-free RNA before RT-PCR and RT-qPCR reactions;combine with DNA polymerase I to perform DNA labelingthrough nick translocation;DNA fragmentation library construction.

This product is produced in accordance with GMP process requirements,and the product is provided in liquid form.

Components

Storage conditions

This product should be stored at-25~- 15℃ for 2 years. Please avoid repeated freeze thaw.

Specifications

Instructions

1.Plasmid  template  digestion

1.1 Reaction system:

Use the RNase-free centrifuge tube and pipette tip to prepare the following reaction system:

*10×DNase  IBuffer:10  mM Tris-HCl,2.5mM  MgClb,0.5mMCaClz,pH7.6at25℃

1.2 Reaction conditions

37℃,15-30min  later,add  afinal  concentration  of2.5mMEDTAsolutionand  mixwellat65℃for  10min.The  processed template can be used in subsequent reactions such as capping reaction

2.DNase      linactivationor      inhibition

After adding EDTA to a final concentration of 2.5mM,heating at 65℃for 10 min can inactivate DNase I.Phenol and chloroform extraction can also inactivate DNase l.The following conditions all have significant inhibitory effect on DNase   l:Metal   ion   chelating   agents,zinc   ions   with    a    concentration    of    millimoles/liter,0.1%SDS,DTT, mercaptoethanol and other reducingagents,the salt concentrations above 50-100 mM.

Notes

1.Enzymes should be stored in an icebox or on an ice bath when used,and should be stored at-20℃immediately after use.

2.For  your  safety  and  health,please  wear  personal  protective  equipment(PPE),such  as  laboratory  coats  and disposable gloves,when operating with this product.