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Chomera tsamba Direct PCR kuphatikiza zida-UNG (popanda Zitsanzo Zida) Protocol Direct PCR from Plant Material

Kufotokozera Kwa Zida:

Mwachindunji ntchito polysaccharide ndi polyphenol tsamba lysate monga Chinsinsi PCR matalikidwe, popanda osiyana m'zigawo za DNA, kudya ndi mkulu tilinazo.Dongosolo la UNG loletsa kuwononga chilengedwe lawonjezedwa kuti tipewe zolakwika.

Palibe kuyeretsa kwa DNA kowononga nthawi komanso kokwera mtengo (Nthawi, ntchito ndi mtengo wasungidwa!)

Zinthu zochepa

Zosavuta-chitsanzocho chikhoza kuchitidwa ndi PCR kapena lysis reaction popanda kudula kapena kugaya

2 X Mix-kuchepetsa zolakwika pakukweza zitsanzo ndi nthawi yokonzekera dongosolo

Kukonzekera mwachangu-template kumatha kumaliza mphindi 10, machitidwe a PCR amatha kumaliza mphindi 50

High throughput-lysis reaction itha kumalizidwa mu mbale ya 96-PCR

mphamvu zam'tsogolo


Tsatanetsatane wa Zamalonda

Zolemba Zamalonda

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

Plant Leaf Direct PCR Plus Kit-UNG4

Electropherogram

Plant Leaf Direct PCR Plus Kit-UNG5

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

Plant Leaf Direct PCR Plus Kit-UNG4

Electropherogram

Plant Leaf Direct PCR Plus Kit-UNG5

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