Methylation-specipc PCR (MSP) Kit

Methylation-yakatarwa PCR yekuona kit.

Methylation-speci fi c PCR (MSP) Kit yakagadzirirwa zvakanyanya kune vatengi vanoongorora maitiro e methylation e genomic DNA nePCR, umo MSP DNA Polymerase iri thermostable polymerase yakagadziridzwa nema antibodies, uye 10 × MSP PCR Buffer ndeye PCR buffer yakagadzirirwa kunyanya MSP kuita. Inoenderana neTIANGEN DNA Bisulfite Conversion Kit (4992447).

Cat. Aihwa Kurongedza Saizi
4992759 50 preps

Chigadzirwa Chigadzirwa

Kufamba kwebasa

Ongororo Mienzaniso

MIBVUNZO

Chigadzirwa Mateki

Zvimiro

■ Chigadzirwa chine zvakanakira kukurumidza, kureruka, kushushikana kwakanyanya, kusimba kwakasimba uye kudzikama kwakanaka.

Tsanangudzo

Type: MSP DNA Polymerase
Template: <500 ng
Mafomu: Inokodzera methylation yakatarwa PCR (MSP) nzira yekuongorora maitiro e methylation e genomic DNA

Zvese zvigadzirwa zvinogona kuve zvakagadziriswa ODM / OEM. Kuti uwane mamwe mashoko,ndokumbira ubaye Customized Service (ODM / OEM)


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    1. Methylation-Yakatsanangurwa PCR (MSP) Kit yakashandiswa kuwedzera 400 bp chidimbu uchishandisa bisulfite yakarapwa genomic DNA setemplate ine reaction system ye20 μl.

    Experimental Exampl

    2. Kugadzwa kwePCR reaction kutenderera

    Experimental Exampl

     

    Experimental Examples 1: Samples inogadziriswa neMutengesi A;
    2: Samples inogadziriswa neMutengesi B;
    3: TIANGEN yakarapwa sampuro 4: NTC; M: D2000
    Experimental Examples 1-6:
    6 inodzokorora;
    7: Mutengesi Muenzaniso wakagadziriswa;
    8: NTC; Bio2-F / R uye P16- Me-F / R: Maviri ekuona ekutanga.
    Mubvunzo: Hapana mabhendi ekusimudzira

    A-1 Chidimbu

    ■ Iyi template ine mapuroteni marara kana kuti Taq inhibitors, nezvimwe- ——Chenesa DNA template, bvisa tsvina yeprotein kana bvisa template yeDNA nemidziyo yekuchenesa.

    ■ Kuburitswa kwetemplate hakuna kukwana - - Zvakakodzera kuwedzera tembiricha yedenature uye kuwedzera nguva yekushandurwa.

    ■ Kushatiswa kwetemplate ——Gadzirisazve template.

    A-2 Primer

    ■ Hunhu hwemhando dzekutanga - - Re-gadzira iyo yekutanga.

    Prim Kushatiswa kwekutanga --—Rovera mabara epamusoro epamusoro kuva vhoriyamu diki yekuchengetedza. Dzivisa kuwanda kwechando uye kunyungudika kana kwenguva refu -4 ° C yakachengetedzwa.

    ■ Dhizaini isiriyo yezvigadzirwa (semuenzaniso, urefu hwepamberi haukwanire, dhayera yakaumbwa pakati pezvakagadzirwa, nezvimwewo) -Reresign primers (dzivisa kuumbwa kweprimer dimer uye yechipiri chimiro)

    A-3 Mg2+kuisa pfungwa

    ■ Mg2+ kuiswa kwakanyanya kwazvo ——Kuwedzera zvakafanira Mg2+ kusangana: Gonesa iyo Mg2+ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg2+ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

    A-4 Kuwedzera kwekushisa

    The Iyo tembiricha yekukwirisa inokanganisa kusunga kwekutanga uye template. - -Deredza iyo yekuwedzera tembiricha uye gadzirisa mamiriro acho ne gradient ye2 ° C.

    A-5 Yekuwedzera nguva

    ■ Pfupi yekuwedzeredza nguva - - Wedzera nguva yekuwedzera.

    Mubvunzo: Nhema dzakanaka

    Phenomena: Negative sampuli zvakare inoratidza iwo akanangana akateedzana mabhendi.

    A-1 Kusvibisa PCR

    ■ kusvibiswa kwemuchinjikwa kwekutevedzana kwekutarisa kana zvigadzirwa zvekusimudzira - - Nekuchenjera kusapomba sampuro ine nhevedzano yechinangwa mune yakashata sampuro kana kuvadururira kunze kwecentrifuge chubhu. Iwo ma reagents kana zvishandiso zvinofanirwa kuve zvakagadziriswa kuti zvibvise zviripo nucleic acids, uye kuvepo kwehutachiona kunofanirwa kutemerwa kuburikidza neasina kudzora kuyedza.

    ■ Kusvibisa reagent ——Root the reagents and store at low low.

    A-2 Primer

    ■ Mg2+ kuiswa kwakanyanya kwazvo ——Kuwedzera zvakafanira Mg2+ kusangana: Gonesa iyo Mg2+ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg2+ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

    ■ Dhizaini isina kukodzera, uye maratidziro akanangwa ane humology pamwe neasingaitane nhevedzano. —-Re-dhizaini yekutanga.

    Mubvunzo: Asina-chaiwo kududzira

    Phenomena: Iwo maPC ekukudziridza mabhendi haapindirane nehukuru hunotarisirwa, ingave hombe kana diki, kana dzimwe nguva zvese zvakasarudzika mabhendi ekuwedzera uye asiri-akasarudzika mabhendi ekusimudzira anoitika.

    A-1 Primer

    ■ Kusanyatsojeka kwekutanga

    —-Re-dhizaini yekutanga.

    ■ Iko kusungirwa kwepamusorosoro kwakanyanyisa - - Zvakakodzera kuwedzera tembiricha yedenature uye kuwedzeredza nguva yekushanduka.

    A-2 Mg2+ kuisa pfungwa

    ■ Iye Mg2+ kusungwa kwakanyanya ——Deredza zvakanaka iyo Mg2 + kukosheswa: Gonesa iyo Mg2+ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg2+ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

    A-3 Thermostable polymerase

    ■ Huwandu hwe enzyme yakawandisa ——Deredza huwandu hwee enzyme nenzira kwayo panguva dze 0.5 U.

    A-4 Kuwedzera kwekushisa

    ■ Tembiricha yekuvharisa yakanyanya kushomeka - - Zvakakodzera kuwedzera tembiricha yekuwedzera kana kutora nzira mbiri yekuvharisa

    A-5 PCR kutenderera

    ■ Makwikwi mazhinji ePDR ——Deredza huwandu hwePCR macircule.

    Mubvunzo: Patchy kana smear mabhendi

    A-1 Primer-—Kusarongeka kwehurombo--- Gadzirisazve iyo yekutanga, chinja chinzvimbo uye kureba kweiyo yekutanga kuti iwedzere kujekesa kwayo; kana kuita PCR nested.

    A-2 template DNA

    ——Template haina kuchena— - Chenesa template kana bvisa DNA nemidziyo yekuchenesa.

    A-3 Mg2+ kuisa pfungwa

    ——Mg2+ kuisa pfungwa kwakanyanya ——Deredza zvakanaka Mg2+ kusangana: Gonesa iyo Mg2+ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg2+ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

    A-4 dNTP

    ——Kusungwa kweDNTPs kwakanyanyisa ——Deredza kusangana kweDNTP nenzira kwayo

    A-5 Kuwedzera tembiricha

    ——Tembiricha yakadzikira chose inovhara —— Zvishoma nemazvo wedzera tembiricha yekuwedzera

    A-6 Matunhu

    --—Kwakawanda kutenderera —— Wedzera huwandu hwenhamba

    Mubvunzo: Mari yakawanda sei template inofanirwa kuwedzerwa mune 50 μl PCR reaction system?
    ytry
    Mubvunzo: Ungawedzera sei zvidimbu zvakareba?

    Nhanho yekutanga ndeyekusarudza yakakodzera polymerase. Yakajairwa Taq polymerase haigone kuongorora nekuda kwekushayikwa kwe3'-5 'exonuclease chiitiko, uye kusaenderana kuchadzora zvakanyanya kuwedzera kwekuwedzera kwezvimedu. Naizvozvo, yenguva dzose Taq polymerase haigone kunyatsokwenenzvera zvidimbu zvakanangwa zvakakura kupfuura 5 kb. Taq polymerase ine yakakosha nadzurudzo kana imwe yakanyanya kutendeka polymerase inofanirwa kusarudzwa kuvandudza kuwedzera kwekuita uye kuzadzikisa zvido zvehurefu hwakareba kuwedzera. Uye zvakare, kukwidziridzwa kwezvimedu zvidimbu kunodawo kuenderana kunoenderana kwekutanga dhizaini, nguva yekuwedzeredza, nguva yekuwedzera, buffer pH, nezvimwe. Kazhinji, ma primers ane 18-24 bp anogona kutungamira mukukohwa kuri nani. Kuti tidzivirire kukuvara kwetemplate, iyo nguva yekuratidzira iri pa94 ° C inofanirwa kudzikiswa kusvika pamasekondi makumi matatu kana mashoma pasiringi, uye iyo nguva yekukwira tembiricha kusvika pa94 ° C pamberi pekukudziridza inofanira kunge iri pasi pe1 min. Zvakare, kumisikidza iyo yekuwedzera tembiricha ingangoita se68 ° C uye nekugadzira iyo yekuwedzera nguva zvinoenderana nehuremu hwe1 kb / min inogona kuve nechokwadi chinoshanda kukwidziridzwa kwezvimedu zvidimbu.

    Mubvunzo: Ungavandudza sei kukwidziridzwa kuvimbika kwePCR?

    Chiyero chekukanganisa kwePCR kukwidziridzwa kunogona kudzikiswa nekushandisa akasiyana maDNA ma polymerase nekuvimbika kwakanyanya. Pakati pese maTaq DNA ma polymerases akawanikwa kusvika parizvino, Pfu enzyme ine yakaderera kukanganisa chikanganiso uye yakatendeka kuvimbika (ona tafura yakasungirirwa). Pamusoro pekusarudzwa kwe enzyme, vatsvagiri vanogona kuwedzera kudzikisa huwandu hwePCR nekushandura mamiriro ekugadzirisa, kusanganisira kugadziridza bhaisikopo, kuwanda kweinopisa polymerase uye kugadzirisa huwandu hwePCR.

    Nyora yako meseji pano uye uitumire kwatiri