Ultra Nuclease GMP-kōeke
Cat Nama: HC2016A
Ko te UltraNuclease GMP-grade e whakaatuhia ana, e purea ana i roto i te Escherichia coli (E.coli) na te hanga ira me te whakarite i raro i nga taiao GMP.Ka taea te whakaiti i te pokey o te supernatant pūtau me te lysate pūtau i roto i te rangahau pūtaiao, te whakanui ake i te pai o te purenga pūmua me te whakarei ake i te rangahau mahi pūmua.Ka taea hoki e te hua te whakaheke i nga toenga waikawa nucleic host ki te tohu-pg, te whakapai ake i te mahi me te haumaru o nga hua koiora o nga tono tae atu ki te purenga huaketo, te hanga kano kano, me te hanga rongoa pūmua/polysaccharide.I tua atu, ka taea hoki te whakamahi i te hua ki te aukati i te kopiri o nga pūtau mononuclear toto peripheral tangata (PBMC) i roto i te rongoa pūtau me te whanaketanga kano.
Ka whakaratohia te UltraNuclease i roto i te ahua o te reagent kua whakahoroa, kua whakahekehia ki roto i te parepare (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), me te ahua o te wai kore tae, maramara.Ko tenei hua ka hangaia e nga whakaritenga tukanga GMP ka tukuna i roto i te ahua wai.
Waehanga
UltraNuclease GMP-kōeke (250 U/μL)
Nga Tikanga Rokiroki
Ka tukuna te hua me te hukapapa maroke ka taea te rongoa ki te -25 ℃ ~ -15 ° C mo nga tau e rua.
Mena ka whakatuwherahia te hua ka penapena ki te 4 ℃ mo te neke atu i te wiki, ka tūtohu kia tātarihiate hua hei aukati i te poke moroiti.
Whakatakotoranga
| Kaihautu Whakaaturanga | Recombinant E. coli me te ira UltraNuclease |
| Taumaha Molecular | 26.5 kDa |
| tohu hiko | 6.85 |
| Maamaa | ≥99% (SDS-WĀRANGI) |
| Pupuri Rokiroki | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Whakamaramatanga Waeine | Ko te whakamaramatanga o tetahi wae Mahi (U) ko te nui o te whākōkī e whakamahia anaHurihia te uara whakauru o te ΔA260 mā te 1.0 i roto i te 30 meneti i roto i te 2. 625 mLpūnaha tauhohenga i te 37 ℃ me te pH o 8.0 (he rite ki te whakaoti nakunaku o37 μg DNA parāoa hāmana ki oligonucleotides). |
Tohutohu
1. Tauira Kohinga
Pūtau piri: tango i te reo, horoia nga pūtau ki te PBS, ka tango i te supernatant.
Ngā pūtau whakatārewatanga: kohia ngā pūtau mā te centrifugation, horoia ngā pūtau ki te PBS, centrifuge i te 6,000rpm mo te 10 meneti, kohia te pire.
Escherichia coli: kohia nga huakita ma te centrifugation, horoi kotahi ki te PBS, centrifuge i te 8,000rpm mo te 5 meneti, ka kohia te pire.
2. Tauira Maimoatanga
Tuhia nga pire pūtau kua kohia me te lysis buffer i te ōwehenga o te papatipu(g) ki te rōrahi(mL)1:(10-20), ma te mahi miihini, matū ranei i runga i te hukapapa, i te pāmahana rūma ranei (1g o te pellet pūtau kei roto
109 pūtau).
3. Te maimoatanga Enzyme
Tāpirihia te 1-5mM MgCl ki te pūnaha tauhohenga me te whakatika i te pH ki te 8-9.
Tāpirihia te UltraNuclease kia rite ki te ōwehenga o te 250 Waeine ki te keri i te 1 g o nga pire pūtau, whakakororia i te 37 ℃ neke atu i te 30 meneti.Tena koa tirohia te puka "Wā Tauhohenga Manakohia" hei whiriwhiri i tete roanga o te maimoatanga.
4. Hunga Kohinga
Whakanuia ki te 12,000 rpm mo te 30 meneti ka kohia te supernatant.
Tuhipoka: Mena he waikawa, he kawakore ranei te otinga, kei roto ranei he tote teitei, he horoi horoi, hedenaturants, tēnā whakanuia te inenga whākōkī, whakaroahia ranei te wa maimoatanga kia rite.
E taunaki ana nga tikanga tauhohengaons
| Tawhā | Tikanga tino pai | Whakaritenga Whaihua |
| Mg²+Kutanga | 1-5 mM | 1-10 mM |
| pH | 8-9 | 6-10 |
| Te pāmahana | 37℃ | 0-42 ℃ |
| DTT Te Kohanga | 0-100 mM | >0 mM |
| Mercaptoethanol Concentration | 0-100 mM | >0 mM |
| Monovalent Cation Concentration | 0-20 mM | 0-150 mM |
| Phosphate lon Concentration | 0-10 mM | 0-100 mM |
E taunaki ana Tauhohenga Te wa (37℃, 2 mM Mg²+, pH 8.0)
| Te Moni UltraNuclease (Whakamutunga Whakamutunga) | Wā Tauhohenga |
| 0.25 U/mL | >10 h |
| 2.5 U/mL | >4h |
| 25 U/mL | 30 min |
Tuhipoka:
Tena koa mauhia nga PPE e tika ana, he koti taiwhanga me nga karapu, kia pai ai to hauora me to haumaru!
