Chomera tsamba Direct PCR kuphatikiza zida-UNG (popanda Zitsanzo Zida) Protocol Direct PCR from Plant Material
Kufotokozera
Kutengera zida za Plant Leaf Direct PCR Plus, dUTP imagwiritsidwa ntchito m'malo mwa dTTP, ndipo puloteni ya UNG yomwe ingawononge template yomwe ili ndi dUTP imawonjezedwa.Mwanjira iyi, kutsimikizika ndi kulondola kwa kukulitsa kumatsimikizika, ndipo vuto la kuipitsidwa kwazinthu za PCR zomwe zitha kuchitika pakuyesa kwakukulu kwa majini zimatetezedwa.
Zofotokozera
50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns
Zida zamagulu
Gawo I | Mtengo P1 |
Mtengo P2 | |
6×DNA Loading Buffer | |
Gawo II | 2× Leaf PCR EasyTMMix(UNG) |
Mbali & ubwino
■ Palibe kuyeretsa kwa DNA kowononga nthawi komanso kokwera mtengo
■ Zochepa
■ Chosavuta-chitsanzo chikhoza kuchitidwa ndi PCR kapena lysis reaction popanda kudula kapena kugaya
■ 2 × Mix, kuchepetsa chitsanzo Mumakonda zolakwa ndi anachita dongosolo kukonzekera nthawi
■ Kukonzekera mwachangu template kumatha kumaliza mphindi 10, kuchita kwa PCR kumatha kutha mphindi 50.
■ High throughput-lysis reaction akhoza kumalizidwa mu 96-chitsime PCR mbale
Zosintha za Kit
Kugwiritsa ntchito: chizindikiritso chosinthidwa ma genetic, genotyping, etc.
Chitsanzo: tsamba la mbewu
Mlingo: Diameter 2-3mm (njira yolunjika), m'mimba mwake 5-7mm (njira ya lysis)
Mtundu wozindikiridwa: chidutswa chandamale ≤1kb
Kuyenda kwa ntchito
Electropherogram
Zofotokozera
50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns
Zida zamagulu
Gawo I | Mtengo P1 |
Mtengo P2 | |
6×DNA Loading Buffer | |
Gawo II | 2× Leaf PCR EasyTMMix(UNG) |
Mbali & ubwino
■ Palibe kuyeretsa kwa DNA kowononga nthawi komanso kokwera mtengo
■ Zochepa
■ Chosavuta-chitsanzo chikhoza kuchitidwa ndi PCR kapena lysis reaction popanda kudula kapena kugaya
■ 2 × Mix, kuchepetsa chitsanzo Mumakonda zolakwa ndi anachita dongosolo kukonzekera nthawi
■ Kukonzekera mwachangu template kumatha kumaliza mphindi 10, kuchita kwa PCR kumatha kutha mphindi 50.
■ High throughput-lysis reaction akhoza kumalizidwa mu 96-chitsime PCR mbale
Zosintha za Kit
Kugwiritsa ntchito: chizindikiritso chosinthidwa ma genetic, genotyping, etc.
Chitsanzo: tsamba la mbewu
Mlingo: Diameter 2-3mm (njira yolunjika), m'mimba mwake 5-7mm (njira ya lysis)
Mtundu wozindikiridwa: chidutswa chandamale ≤1kb