prou
Alaabta
Sawirka BsaI HCP1014A
  • BsaI HCP1014A

BsaI


Bisadda No:HCP1014A

Xidhmada: 50μL/250μL/1ml/10ml/100ml

BsaI, oo ah IIs xaddidaadda endonuclease xaddidaadda endonuclease, ayaa laga soo qaatay isku-dhafka E.

Sharaxaada Alaabta

Xogta alaabta

BsaI, an IIs restriction endonuclease restriction endonuclease, waxaa laga soo qaatay arecombinant E. coli strain kaas oo xanbaarsan hidda-wadaha BsaI ee cloned iyo wax laga beddelay oo ka yimid Bacillus stearothermophilus. Kuwaas oo leh sifooyin isku mid ah sida enzyme hooyo iyo hoos u dhaca dhaqdhaqaaqa xiddigaha.Siday isugu xigto aqoonsiga iyo goobaha kala-goynta waa sidan soo socota:

5'······GGTCTC(N)·············3'

3'······CCAGAG(NNNN)···5'


  • Hore:
  • Xiga:

  • Qaybaha

    BsaI(20U/μL), 10xBsaI Buffer

     

    Kaydinta

    Ku kaydi at -25℃~-15℃, shaqaynaysa 1 sano (iska ilaali wareegyada qaboojinta ee soo noqnoqda).

     

    Kaydinta kaydinta

    10mM Tris-HCl, 200mM NaCl, 1mM DTT 0.1mM EDTA, 200µg/ml Recombinant Albumin,50% Glycerol.(pH 7.4 @ 25°C).

     

    Tilmaamaha alaabta

    Dhaqdhaqaaq sare, dheefshiidka degdega ah;

    1. Dhaqdhaqaaqa xiddiga oo hooseeya, hubinta goynta saxda ah sida "scalpel";

    2. Bilaash la'aanta BSA iyo xoolaha asalkoodu yahay.

    Dareenka Methylation

    Methylation-ka biyo-xireenka: Ma aha xasaasi;

    Methylation-ka DCm: Wax u dhimay Qaar ka mid ah Isku-darka Isku-darka;

    Methylation CpG: Waxaa xannibay Qaar ka mid ah Isku-dhafka Isku-dhafan.

     

     

    Qeexitaan Unug

    Hal mid oo aan la qeexin ayaa lagu qiyaasaa qiyaasta insaymiska loo baahan yahay si loo dheefshiido 1 µg ee kontoroolka gudaha DNA 1 saac 37°C wadarta mugga falcelinta ee 50 µL.

     

    Xakamaynta tayada

    Qiimaynta Nadiifinta Barootiinka (SDS-PAGE)Nadiifinta Bsa I waxay ahayd ≥ 95% waxaa go'aamiyay falanqaynta SDS-PAGE iyadoo la adeegsanayo ogaanshaha Coomassie Blue.

    RNase:20 U of Bsa I oo leh 1.6 μg MS2 RNA oo loogu talagalay 16hoursat37℃ wax-soosaarnodegradaationas waxaa go'aamiyay agarose gel electrophoresis.

    Waxqabadka DNA-ga Aan Gaarka ahayn:20 Uof BsaI oo leh 1 μg PhiX174 DNA 16 saacadood oo ah 37 ℃ ma keeno DNA xad dhaaf ah sida lagu go'aamiyay agarose gel electrophoresis.

    Dib u dhigista iyo dib u dhigista:Ka dib dheefshiidka 1 μg

    E.coli DNA:2Uof VacciniavirusCapping Enzyme waxaa lagu baara joogitaanka E. coli genomic DNA iyadoo la isticmaalayo TaqManqPCR oo leh qalabyo u gaar ah E. coli 16S rRNA goobta.E. coli genomic DNA-da wasakhaysan waa ≤1 E. coli genome.

    Bakteeriyada Endotoxin:Imtixaanka LAL, marka loo eego daabacaadda ChinesePharmacopoeiaIV2020, habka tijaabada xadka jel, qaanuunka guud (1143).Bakteeriyada endotoxin waa inay noqotaa ≤10 EU/mg.

     

    Nidaamka falcelinta iyo xaaladaha

    Qayb

    Mugga

    BsaI (20 U/μL)

    1μL

    DNA

    1μg

    10 x BsaI Buffer

    5μL

    dd H2O

    Ilaa 50 μl

    Dubo at 37 ° C 15-30 daqiiqo.Dhaqdhaqaaq la'aanta kulaylka:80°C 20 daqiiqo

    Nidaamka falcelinta lagu taliyay iyo xaaladaha ayaa ku siin kara saameyn dheefshiidka enzyme aad u wanaagsan, taas oo ah tixraac kaliya, fadlan tixraac natiijooyinka tijaabada ah ee faahfaahinta.

     

    Codsiyada

    Xannibaadda Enzyme Dheefshiidka Xakamaynta Degdegga ah.

     

    Qoraalada isticmaalka

    1.Mugga enzyme ≤ 1 / 10 ee mugga falcelinta.

    2. Dhaqdhaqaaqa-star-ka-shaqsiyeedka waxa uu dhacaa marka fiirsashada glycerol ay ka badan tahay 5%.

    3. Dhaqdhaqaaqa nadiifinta ayaa dhici karta marka substrate ka hooseeyo saamiga lagu taliyey.

     

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