Rnase Inhibitor (Glycerol free)
ʻO Murine RNase inhibitor kahi mea hoʻopaneʻe murine RNase hou i hōʻike ʻia a hoʻomaʻemaʻe ʻia mai E.coli.Hoʻopaʻa ia iā RNase A, B a i ʻole C ma kahi ratio 1: 1 ma o ka hoʻopaʻa ʻana non-covalent, no laila ke keʻakeʻa nei i ka hana o nā enzyme ʻekolu a pale i ka RNA mai ka hoʻohaʻahaʻa ʻana.Eia nō naʻe, ʻaʻole kūpono ia i ka RNase 1, RNase T1, S1 Nuclease, RNase H a i ʻole RNase mai Aspergillus.Ua hoʻāʻo ʻia ʻo Murine RNase inhibitor e RT-PCR, RT-qPCR a me IVT mRNA, a ua kūpono me nā ʻano transcriptases Reverse pāʻoihana, DNA polymerases a me RNA polymerases.
Ke hoʻohālikelike ʻia me nā mea hoʻopaneʻe RNase kanaka, ʻaʻole i loaʻa i ka inhibitor RNase murine ʻelua mau cysteine i koʻikoʻi loa i ka oxidation e hoʻoulu ai i ka inactivation o ka inhibitor.ʻO ia ka mea e paʻa ai i nā haʻahaʻa haʻahaʻa o DTT (emi ma mua o 1 mM).He kūpono kēia hiʻohiʻona no ka hoʻohana ʻana i nā hopena kahi i kū ʻole ai ka nui o ka DTT i ka hopena (e laʻa me RT-PCR Real-time).
Application
Hiki ke hoʻohana nui ʻia kēia huahana i kēlā me kēia hoʻokolohua kahi hiki ke hoʻopili ʻia ʻo RNase e pale aku i ka hōʻino ʻana o RNA, e like me:
1.First-strand cDNA synthesis, RT-PCR, RT-qPCR, etc.;
2. E pale i ka RNA mai ka ho'oha'aha'a 'ana i loko o ke kope in-vitro/unuhi (e la'a me ka ho'opi'i viral in vitro);
3.Inhibition o RNase hana i ka wā RNA kaawale a me ka hoʻomaʻemaʻe.
Nā Kūlana Waihona
E kūʻai ma -25～-15 ℃;
Nā pōʻai hoʻoheheʻe hoʻoheheʻe ≤ 5 mau manawa;
Pono no 1 makahiki.
Wehewehe ʻāpana
Hoʻokahi ʻāpana i wehewehe ʻia e like me ka nui o ka RNase Inhibitor e koi ʻia e kāohi i ka hana o 5 ng o RNase A e 50%.
Kaumaha molekula
ʻO RNase Inhibitor (Glycerol-free) he protein 50 kDa.
Ka hoomalu maikai
Exonuclease Hana:
ʻO ka incubation o 40 U o ka enzyme me 1 μg λ-Hind III digest DNA no nā hola 16 ma 37 ° C ka hopena i ʻike ʻole ʻia ka degradation o ka DNA e like me ka hoʻoholo ʻana e ka electrophoresis gel.
Endonuclease hana:
ʻO ka incubation o 40 U o ka enzyme me 1 μg λ DNA no 16 mau hola ma 37 ° C ka hopena i ʻike ʻole ʻia ka degradation o ka DNA e like me ka hoʻoholo ʻana e ka electrophoresis gel.
Nicking Hana:
ʻO ka incubation o 40 U o ka enzyme me 1 μg pBR322 no 16 mau hola ma 37 ℃ ka hopena i ʻike ʻole ʻia ka hōʻino ʻana o ka DNA e like me ka hoʻoholo ʻana e ka electrophoresis gel.
RNase Ka hana:
ʻO ka incubation o 40 U o ka enzyme me 1.6 μg MS2 RNA no 4 mau hola ma 37 ℃ ka hopena ʻaʻole i ʻike ʻia ka degradation o ka RNA e like me ka hoʻoholo ʻana e ka electrophoresis gel.
E.coli DNA:
ʻIke ʻia ʻo 40 U o Enzyme e TaqMan qPCR.ʻO ka DNA E.coli he ≤ 0. 1pg/40U.
Notes
1. Mai lulu a hoʻoulu ikaika e pale i ka enzyme inactivation.
2. RNase inhibitor mea hana ma ka wela mai 25 ℃ a hiki i 55 ℃ a inactivated ma ≥65 ℃.
3.ʻAʻole i kāohiʻia ka hana a RNase H, RNase 1, a me RNase T1.
4. He ākea ka pae pH no ka pale ʻana i ka hana RNase (ʻeleʻele i ka pH 5-9), e hōʻike ana i ka hana kiʻekiʻe ma pH 7-8.
