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Iqabunga lesitshalo Direct PCR kit-UNG (ngaphandle kwamathuluzi okusampula) Iphrothokholi eqondile ye-PCR evela ku-Plant Material

Incazelo yekhithi:

Sebenzisa ngokuqondile iqabunga noma i-lysate yeqabunga njengesifanekiso sokukhulisa i-PCR, ngaphandle kokukhipha okuhlukile kwe-DNA, ukuzwela okusheshayo nokuphezulu.Uhlelo lwe-UNG lokulwa nokungcoliswa kwemvelo lwengezwe ukuze kuvinjelwe imibono engamanga.

Akukho ukuhlanzwa kwe-DNA okudla isikhathi nokubizayo (Isikhathi, umsebenzi nezindleko zilondoloziwe!)

Impahla encane

Kulula-isampula ingaba ngaphansi kokusabela kwe-PCR noma ukusabela kwe-lysis ngaphandle kokusika noma ukugaya

2 X Mix—ukunciphisa amaphutha okulayisha amasampula kanye nesikhathi sokulungiselela isistimu yokusabela

Ukulungiselela isifanekiso esisheshayo kungaqedwa emizuzwini eyi-10, ukusabela kwe-PCR kungaqedwa emizuzwini engama-50

Ukusabela okuphezulu kwe-throughput—lysis reaction kungaqedelwa kupuleti le-PCR elinemithombo engu-96

amandla angaphambili


Imininingwane Yomkhiqizo

Omaka bomkhiqizo

Izincazelo

Ngokusekelwe kukhithi ye-Plant Leaf Direct PCR, i-dUTP isetshenziswa esikhundleni se-dTTP, futhi i-enzyme ye-UNG engehlisa isithunzi isifanekiso esiqukethe i-dUTP.Ngale ndlela, ukucaciswa nokunemba kokukhuliswa kuyaqinisekiswa, futhi inkinga yokungcoliswa komkhiqizo we-PCR okungenzeka ngesikhathi sokuhlolwa kofuzo okukhulu kuyavinjelwa.

Imininingwane

50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns

Izingxenye zekhithi

Ingxenye I

Isilondolozi se-P1

Isilondolozi se-P2

6×I-DNA Loading Buffer

Ingxenye II

2× Leaf PCR EasyTMImiksi(UNG)

Izici&izinzuzo

■ Akukho ukuhlanzwa kwe-DNA okuchitha isikhathi nokubizayo
■ Impahla encane
■ Kulula-isampula ingaba ngaphansi kokusabela kwe-PCR noma ukusabela kwe-lysis ngaphandle kokusika noma ukugaya
■ 2 × Mix, ukunciphisa amaphutha okulayisha amasampula kanye nesikhathi sokulungiselela uhlelo lokusabela
■ Ukulungiselela isifanekiso esisheshayo kungaqedwa ngemizuzu eyi-10, ukusabela kwe-PCR kungaqedwa okungenani ngemizuzu engama-50
■ Ukusabela okuphezulu kwe-throughput-lysis kungaqedwa ngepuleti le-PCR le-96-well

Imingcele yekhithi

Isicelo: ukuhlonza okushintshwe izakhi, i-genotyping, njll.
Isampula: iqabunga lesitshalo
Isikali: Ububanzi 2-3mm (indlela eqondile), ububanzi 5-7mm (indlela ye-lysis)
Ibanga lokutholwa: isiqeshana esiqondiwe ≤1kb

Ukugeleza komsebenzi

I-Plant Leaf Direct PCR Kit-UNG4

Electropherogram

I-Plant Leaf Direct PCR Kit-UNG5

Amaviki amabili emiphumela yokutholwa kwe-PCR Mix ngaphansi kwesimo sokungcoliswa komkhiqizo wokukhulisa i-PCR.


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