Chida Cha HRM Analysis (EvaGreen)

Mawonekedwe

■ Kusintha kwakukulu: Landirani utoto wonenepa wa EvaGreen, womwe umatha kusungunuka bwino kwambiri munthawi yodzaza ndipo umatha kusiyanitsa kusiyanasiyana kwa maziko amodzi.
■ Kutsogola kwapamwamba: Gwiritsani ntchito DNA polymerase yoteteza ku anti-anti-anti-anti-amplification kuti muchepetse kukulitsa komanso kusasintha.
■ Kukhazikika kwakukulu: Makina ogwiritsira ntchito mosamala bwino amakulitsa kukhazikika kwa kukhazikika ndi kukonza kudalirika kwa zotsatira.
Kukonzekera kwa ROX: Utoto wa ROX umaphatikizidwa padera, womwe umakhala wosavuta kugwiritsa ntchito ndipo umakhala ndi zotsatira zolondola.

Mfundo

Mtundu: Antibody yosinthidwa Taq DNA polymerase, EvaGreen.
Mapulogalamu: Kudziwika kwa SNP; Kuwonetsa SNP kosadziwika; Kusanthula majini osadziwika; Kusanthula kwa Methylation PCR.

Zogulitsa zonse zimatha kusinthidwa kukhala ODM / OEM. Zambiri,chonde dinani Makonda Service (ODM / OEM)

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Genomic DNA idatengedwa kuchokera ku 100 μl yamagazi amunthu pogwiritsa ntchito TIANamp Blood DNA Kit. 50 ng DNA idakwezedwa pompopompo pakuzindikira kwa PCR. Malinga ndi library ya NCBI SNP, mtundu wodziwika wa SNP wa FEN1 gene (RS 174538) umasankhidwa ndikuwonekera pogwiritsa ntchito Roche LightCycle480. Zotsatira zake ndi izi:

Zotsatirazo zikuwonetsa kuti HRM Analysis Kit ndi njira yabwino yosanthula malo a SNP ndikubwereza mobwerezabwereza kosungunuka kwa mtundu womwewo, kuwunika momveka bwino kwamitundu yosiyanasiyana komanso kuweruza molakwika.

Zambiri za SNP: …… CGGAAGAACACGTCG [A / G] CAGGAGCAGGCGCCT …… (Nthawi Zonse: A = 0.314, G = 0.686)

Choyamba: Kwa chidutswa cha bp 108 (FEN1_F: CCTCAACGCTCTCACCATTTTG; FEN1_R: GGCACTTCCTTTTCCGGTTGTG)