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T7 RNA Polymerase HCP1011A Kiʻi Manaʻo
  • T7 RNA Polymerase HCP1011A

T7 RNA Polymerase


Helu popoki:HCP1011A

Pūʻolo: 100μL / 1mL / 10mL / 100mL

ʻO T7 RNA Polymerase kahi RNA polymerase hilinaʻi DNA mai ka T7 phage

Hōʻike huahana

ʻikepili huahana

ʻO T7 RNA Polymerase he RNA polymerase pili i ka DNA mai ka T7 phage, nona ka hana 5'→ 3' RNA polymerase ikaika a kikoʻī. He kikoʻī kiʻekiʻe ka T7 RNA Polymerase no nā kaʻina hoʻolaha T7 a e hoʻohui i ka nui o ka RNA mai kahi ʻāpana DNA. hoʻokomo ʻia i lalo mai kahi mea hoʻolaha.


  • Mua:
  • Aʻe:

  • Nā ʻāpana

    T7 RNA polymerase (50 U/μL)

    10×HH T7 Hoʻopaʻa

    Nā kūlana mālama

    ʻO ka halihali ma lalo o 0°C a me ka waiho ʻana ma -25 ~ - 15 °C.

    Hōʻike

    inoa mea kūʻai

    T7 RNA Polymerase

    Huahana hua

    wai māʻemaʻe

     

    Ka wehewehena hana

    Hoʻokahi ʻāpana i wehewehe ʻia ka nui o ka enzyme

    e hoʻopili i ka NTP e hana i 1 nmol PPi i loko

    1 hola ma 37 ° C ma lalo o ka ʻōnaehana pane maʻamau.

    Paʻa mālama

    50 mM Tris-HCl, 100 mM NaCl, 20 mM β-M, 1 mM EDTA, 50% Glycerol ,0.1% (w/v)

    Triton® X- 100, pH 7.9 @ ​​25°C.

     

    10×HH T7 pale

    400 mM Tris-HCl (25 ℃ , pH 8.20), 60 mM

    MgCl2 ,100 mM DTT ,20 mM spermidine.

    Kūlana mālama

    -25 ~ - 15 ℃ , e pale i ka hoʻolulu hou ʻana

     

    Pane a me ke kūlana

    Pane maʻamau

    Reagent

    Ka nui

    ʻO ka wai i mālama ʻia e ka Nuclease-free H2O a i ʻole DEPC

    A hiki i 20 μL

    10×HH T7 Hoʻopaʻa

    2 μL

    ATP/GTP/CTP/UTP (100 mM kēlā me kēia)

    0.4 μL kēlā me kēia (2 mMeach Hope)

    RNase Inhibitor (40 U/μL) (koho)

    1 μL (40 U)

    Pyrophosphatase Inorganic (koho)

    0.5 μL (0.05U)

    T7 RNA polymerase(50 U/μL)

    1 μL

    Laina DNA Linearized

    1 μg

    Hoʻohui i nā ʻāpana hoʻololi i ke kauoha i luna * 10 × HH T7 Buffer kūpono wale no 2mM NTPs, ʻae ʻia nā kime transcription kiʻekiʻe T7 no 7.5mM-10mM NTPs.

    Manawa Incubation:37 °C no 2 mau hola.

    ʻO Stop o ka pane: Hoʻohui i 2μL 0.2 M EDTA (pH8.0@25 ℃) a i ʻole wela i 75 °C no 10min.

    Hoʻopau DNA: Hiki ke hoʻoneʻe ʻia ka laʻana DNA me 2U DNase I (RNase-free) a me ka incubation no 15 min ma 37 °.

     

    Ka hoomalu maikai

    Endonuclease Activity: Incubation o ka hopena 50μL i loaʻa ka liʻiliʻi o 50U o T7 RNAPolymerase me 1 μg λDNA no 16 mau hola ma 37 ℃ hopena ʻaʻole i ʻike ʻia ka hōʻino e like me ka mea i hoʻoholo ʻia.

    Ka hana Exonuclease: Incubation o ka hopena 50μL i loaʻa ka liʻiliʻi loa o 50U o T7 RNA Polymerase me 1 μg λ -Hind Ⅲ digest DNA no 16 mau hola ma 37 ℃ hopena ʻaʻole i ʻike ʻia ka degradation e like me ka mea i hoʻoholo ʻia.

    Nickase Ka hana: Incubation o ka hopena 50μL i loaʻa ka liʻiliʻi o 50U o T7 RNA Polymerase me 1μg pBR322 DNA no 16 mau hola ma 37°C ʻaʻohe hopena i ʻike ʻia e like me ka mea i hoʻoholo ʻia.

    RNase Ka hana: Incubation o ka hopena 50μL i loaʻa ka liʻiliʻi o 50U o T7 RNA Polymerase me 1.6μg MS2 RNA no 16 mau hola ma 37°C i ka hopena ʻaʻohe degradable ʻike ʻia e like me ka mea i hoʻoholo ʻia.

    ʻAʻohe wela: 75 °C no 10 min.

     

    Mālama

    • Pono e hana 'ia ka hana unuhi ma kahi kaiapuni me ka poke ole RNase.Pono ke komo lima lima.ʻO nā ʻōlelo aʻoaʻo, nā paipu a me ka wai pono ʻole ka nuclease free.

    • Hiki ke hoʻonui i ka hua o ka RNA ma o ka hoʻonuiʻana i ka nui o ka NTP (hiki i kēlā me kēiaʻike ke piʻi i ka 10mM), aʻo ka Mg2+ e pono ke hoʻonui pono.

    • Pono e hoʻomākaukau ʻia ka hui ʻana o ka RNA synthesis reaction ma ka wela lumi, no ka mea, hiki ke hoʻoheheʻe ʻia ka DNA ma ke alo o 10×HH T7 Buffer ma 4°C.

    • E emi ana ka hua o ka loa'a pono 'ana ke ho'onohonoho pono 'ole ka DNA template.• Hiki ke hoʻonui a hoʻohaʻahaʻa i ka hui ʻana.• Inā ho'ēmiʻia ka hopena o ka huahana, hiki ke hoʻohuiʻia he 20 mM DTT hou i ka pūnaewele hoʻonā.

    • ʻO nā ʻāpana i kākau ʻia ʻaʻole i emi a like paha me 500bp, a pono e hoʻolōʻihi i ka manawa kākau i 4-8 mau hola.

    • He maʻalahi ka loaʻa ʻana o nā ʻeke ma 10×HH T7.

    E kākau i kāu leka ma aneʻi a hoʻouna mai iā mākou