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ʻO Fast Ampli RT-qPCR Premix me-UNG HCB5144E Kiʻi Manaʻo
  • Fast Ampli RT-qPCR Premix me-UNG HCB5144E
  • Fast Ampli RT-qPCR Premix me-UNG HCB5144E

ʻO ka wikiwiki Ampli RT-qPCR Premix me-UNG


Helu popoki: HCB5144E

Pūʻolo: 100RXN/1000RXN/10000RXN

Antibody-hoʻololi wela hoʻomaka enzyme, 95 ° C, 1-5min hoʻomaka wela

ʻIke ʻia ʻo qPCR multiplexed multi-channel o nā pahuhopu like ʻole

ʻO ka hoʻonui ʻana i ka naʻau kiʻekiʻe o nā kumu hoʻohālikelike RNA haʻahaʻa

Hoʻonui wikiwiki

Hōʻike huahana

kikoʻī huahana

Helu popoki: HCB5144E

Hoʻokumu ʻia ʻo Fast Ampli RT-qPCR Premix plus-UNG no nā kaʻina hana lyophilization, i noi ʻia no ka nui o ka fluorescence (TaqMan probe) RNA amplification detection i loaʻa ka neocript reverse transcriptase a me ka hoʻonui wikiwiki ʻana i ka DNA Polymerase i loaʻa e ka genetically modified a me ka screening hiki ke hoʻopau i ka PCR amplification i loko o 20 -40 minuke.Hoʻokumu kēia reagent i ka hoʻomanawanui kiʻekiʻe, ʻoi aku ka kiʻekiʻe o ka hoʻohuli ʻana a me ka PCR amplification pono, kūpono no ka hoʻonui ʻana i ka ʻike kiʻekiʻe o nā laʻana RNA haʻahaʻa.Hiki ke loaʻa i kahi pihi maʻamau maikaʻi i loko o kahi ākea quantitative, a hiki ke hoʻokō pololei ʻia ka quantification.Hoʻohana kēia reagent i kahi enzyme hui ʻia o ka enzyme amplification anti-inhibition a me ka enzyme UNG, a me kahi ʻōnaehana buffer i hoʻopaʻa ʻia me ka dUTP.ʻAʻole hiki iā ia ke hoʻokō i ka hoʻonui maikaʻi ʻana o ka gene target i loko o nā laʻana i loaʻa nā inhibitors, akā pale maikaʻi nō hoʻi i ka hoʻonui ʻana i ka maikaʻi hoʻopunipuni i hana ʻia e ka PCR koena a me ka pollution aerosol.Ua kūpono kēia reagent me ka hapa nui o nā mea hana PCR fluorescent quantitative, e like me Applied Biosystems, Eppendorf, Bio-Rad, Roche a pēlā aku.Hiki ke hoʻohana ʻia kēia reagent no ka lyophilization e loaʻa ai kahi ʻano lyophilized maikaʻi a me ke kūpaʻa huahana.


  • Mua:
  • Aʻe:

  • ʻO ka hoʻohui ʻana

    1. 12.5×FastAmpli Part Taq/UNG Mix (me nā dNTP) (DG)

    2. 50× FastAmpli Mahele RTase (DG)

    3. 5 ×FastAmpliRT Buffer (DG)

    4. 4 ×lyoprotectant (koho) (DG)

     

    Nā Kūlana Waihona

    Hiki ke mālama ʻia ma -20 ℃ no ka wā lōʻihi, ma 4 ℃ a hiki i 3 mahina.Hoʻohui maikaʻi ma mua o ka hoʻohana ʻana a palehoʻoheheʻe pinepine.

     

    Kūkākūkā Kaʻikala

     

    ʻanuʻu

     

    Mahana

    PCR mau Kaʻina hana

    PCR wikiwikiKaʻina hana

     

    Nā pōʻaiapuni

    Ka lōʻihi

    Ka lōʻihi

    HuliKākau unuhi

    50 ℃

    15 min

    5 min

    paa

    PolymeraseHo'ola

    95 ℃

    1-5 min

    1-2 min

    paa

    Denature

    95 ℃

    10-20 s

    1-3 sec

    40-50

    Hoʻopili/Hoʻonui

    56-64 ℃

    20-60 s

    3-20 sec

     

    RT-qPCR Liquid ReactPūnaehana ion

    Huina

    25µL Volume

    50µL Volume

    Hoʻopaʻa

    5×FastAmpli RT Buffer

    5µL

    10µL

    12.5×FastAmpli Part Taq/UNG Mix (me nā dNTP)

    2µL

    4µL

    50× FastAmpli Part RTase

    0.5µL

    1µL

    4×lyoprotectant

    6.25µL

    12.5µL

    25×Primer-Probe Mix

    1µL

    2µL

    Kāpena RNA

     

     

     

    ddH2O

    I ka 25µL

    I ka 50µL

    1. He ʻōnaehana lyophilization kēia ʻōnaehana;Ke hoʻohana nā mea kūʻai i kēia ʻōnaehana me ka ʻole o nā koi lyophilization, hiki ke hoʻohui ʻia ka 4 × lyoprotectant;Inā makemake ʻia nā huahana lyophilized, i ka wā o ka wai reagents e hōʻoia i ka hana huahana, pono ia e hoʻohui i ka 4 × lyoprotectant e hōʻoia i ka kūlike me nā ʻōnaehana lyophilized a me nā hopena.

    2. Ke hoʻohana maʻamau PCR kaʻina hana, ka hope loa o ka primer mea maʻamau 0.2μM.No nā hualoaʻa maikaʻi aʻe, hiki ke hoʻopaʻa ʻia ke kumu kumu i loko o ka laulā o 0.2-1.0μM.Hiki ke hoʻopaʻa ʻia ka ʻimi noiʻi i loko o ka laulā o 0.1-0.3μM.Hiki ke hana ʻia nā hoʻokolohua gradient concentration e ʻike i ka hui maikaʻi loa o nā primers a me nā probes.

    3. I ka hoʻohana ʻana i ke ʻano hoʻonui PCR wikiwiki, hiki ke loaʻa kahi hopena maikaʻi aʻe ma o ka hoʻonui ʻana i ka manaʻo o ka primer/probe, a me ka hoʻoponopono ʻana i ka pākēneka o ka primer a me ka probe.

    4. Loaʻa i nā ʻano maʻamau like ʻole ka helu kope o ka gene i manaʻo ʻia.Hiki ke hana i ka dilution gradient no ke koho ʻana i ka nui o ka hoʻohui template kūpono, inā pono.

     

    Hoʻomākaukau ʻōnaehana Lyophilization

    Huina

    25µL Pane Pūnaehana

    5×FastAmpli RT Buffer

    5µL

    12.5×FastAmpli Part Taq/UNG Mix (me nā dNTP) (DG)

    2µL

    50×FastAmpli Part RTase (DG)

    0.5µL

    4×Lyoprotectant

    6.25µL

    25×Primer-Probe Mix

    1µL

    ddH2O

    I ka 18~20µL

    * Inā makemake ʻia nā ʻōnaehana ʻē aʻe no ka lyophilization, e ʻoluʻolu e kūkākūkā kaʻawale.

     

    Lyophilization Process

    Kaʻina hana

    Temp.

    Manawa

    Kūlana

    Paʻi

     Ka hau hau

    4℃

    30 min

    Paʻa

     1 atm

    -50 ℃

    60 min

    Hoʻoluʻu

    -50 ℃

    180 min

    Paʻa

     Hoʻomaloʻo Kumu

    -30 ℃

    60 min

    Hoʻomāhana

     Māmā Loa

    -30 ℃

    720 min

    Paʻa

    Hoʻomaloʻo ʻelua

    25 ℃

    60 min

    Hoʻomāhana

     Māmā Loa

    25 ℃

    300 min

    Paʻa

    1. ʻO kēia kaʻina hana lyophilization he hana hoʻomaloʻo maloʻo in-situ no kahi ʻōnaehana 25µL;inā makemake ʻia nā pahu hoʻomaloʻo maloʻo a i ʻole nā ​​​​kaʻina hana hoʻomaloʻo maloʻo ma loko, e ʻoluʻolu e nīnau kaʻawale.

    2. ʻO ke kaʻina hana lyophilization ma luna nei no ka ʻike wale nō.Loaʻa i nā ʻano huahana like ʻole a me nā mea maloʻo maloʻo like ʻole, no laila hiki ke hoʻoponopono ʻia e like me nā kūlana maoli i ka wā e hoʻohana ai.

    3. He kūpono paha nā kaʻina hana lyophilization like ʻole no nā huahana lyophilized pūʻulu like ʻole, no laila pono e hoʻokō ʻia ka hōʻoia hōʻoia inā hoʻohana ʻia no ka hana nui.

     

    'Ōlelo no ka hoʻohana 'ana i ka lyophilized pauda

    1. Hoʻopili pōkole i ka pauka lyophilized;

    2. E hoʻohui i ka mamana nucleic acid i ka pauka lyophilized a hoʻohui i ka wai a hiki i 25µL;

    3. E hui maikaʻi me ka centrifugation a holo ma ka mīkini.

     

    Ka Mana Mana

    1. Ka ho'āʻo hana: sensitivity, specificity, reproducibility o RT-qPCR.

    2. ʻAʻohe hana nuclease exogenous, ʻaʻohe exogenous endo/exonuclease contamination.

     

    ʻIkepili ʻenehana:

    1. ʻAʻole emi ka nui o ka amplification o ka DNA polymerase wikiwiki ma mua o 1kb/10s.ʻOkoʻa nā mea hana PCR like ʻole me ka hoʻomehana ʻana a me ka wikiwiki hoʻoluʻu, nā ʻano hoʻomalu wela a me nā conductivity thermal, no laila pono ka hoʻonui ʻana i kāu kumu kumu/ʻimi noiʻi a me ke ʻano holo i hui pū ʻia me kāu mea hana PCR wikiwiki.

    2. Hoʻonui ʻia ka manawa unuhi hope e like me ka lōʻihi o ka ʻāpana e hoʻonui ʻia.No nā ʻāpana ʻoi aku ka lōʻihi, hiki ke hoʻolōʻihi ʻia ka manawa kākau hope.

    3. Pono e hoʻonui ʻia ka mahana hoʻonui annealing ma muli o ka waiwai Tm o ka ʻimi mua a me nā kūlana maoli o ka hopena.

    4. No nā mea hoʻomaka me ka mahana annealing haʻahaʻa a ʻoi aku paha ma mua o 200 bp mau ʻāpana, ʻōlelo ʻia ke ʻano 3-step.

    5. E ʻoluʻolu e hoʻohana i nā wahi i hoʻolaʻa ʻia a me nā pipette ma mua a ma hope o ka hoʻonui ʻana, e hoʻokomo i nā mīkina lima a hoʻololi pinepine iā lākou;mai wehe i ka paipu hopena ma hope o ka pau ʻana o ka hopena PCR e hōʻemi i ka hoʻohaumia ʻana o nā laʻana e nā huahana PCR.

    6. ʻOkoʻa ka maikaʻi o ka hoʻohana ʻana o ka dUTP a me ka naʻau i ka enzyme UNG no nā ʻano genes ʻokoʻa, no laila inā hoʻohana ʻia ka ʻōnaehana UNG i ka emi ʻana o ka ʻike maka, pono e hoʻoponopono ʻia ka ʻōnaehana hopena.Inā makemake ʻia ke kākoʻo ʻenehana e ʻoluʻolu e kelepona mai iā mākou.

     

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