Animal Tissue Direct PCR kit-UNG (isina Sampling Zvishandiso) Protocol
Kit Description:
Shandisa zvakananga mhuka yemhuka lysate se template yePCR amplification, pasina kubviswa kwakasiyana kweDNA, kukurumidza uye kunzwisisika kwakanyanya.Yakawedzera UNG anti-kusvibisa sisitimu yekubvisa manyepo.
◮Hapana nguva-inopedza uye inodhura yekuchenesa DNA inodiwa.
◮Sample inodiwa idiki, ingotora mhodzi imwe chete.
◮Hapana mishonga yakakosha yakadai sekugaya nekupwanya inodiwa, uye kushanda kuri nyore.
◮Iyo yakagadziridzwa PCR sisitimu inogonesa PCR kuti ive nemhando yepamusoro uye yakasimba kushivirira kune PCR reaction inhibitors.
Tsanangudzo
Iyi kit inoshandisa yakasarudzika lysis buffer system kukurumidza kuburitsa genomic DNA kubva kumhuka tishu samples kune PCR maitiro, saka inonyanya kukodzera kune yakakura-chiyero genetic kuongororwa.
Iyo nzira yekuburitsa genomic DNA kubva kulysis buffer inopedzwa mukati me10-30 maminetsi pa65 ° C.Hapana mamwe maitiro akadai seprotein uye RNA kubviswa anodiwa, uye yakaburitswa trace DNA inogona kushandiswa setemplate yePCR reaction.
2 × PCR NyoreTMMix (UNG) inoshandisa dUTP pachinzvimbo che dTTP pahwaro hwe2 × PCR EasyTMSanganisa, uye unowedzera UNG enzyme (Uracil-N-glycosylase) inogona kukanganisa template ine dUTP panguva imwe chete.Pamberi pePCR maitiro, iyo UNG enzyme inoshandiswa kusvibisa PCR chigadzirwa chine uracil.Iyo UNG enzyme haizove nemhedzisiro pane template iyo isina uracil, nekudaro ichiva nechokwadi cheiyo chaiyo uye kurongeka kweamplification uye kudzivirira mukana wePCR zvigadzirwa kusvibiswa panguva yakakura-yakakura genetic test.
D-Taq DNA polymerase ndeye DNA polymerase yakanyatso gadzirwa neForegene kune yakananga PCR maitiro.D-Taq DNA polymerase ine kushivirira kwakasimba kune akasiyana PCR reaction inhibitors, uye inokwanisa kunyatso kukwidziridza huwandu hweDNA mune dzakasiyana siyana dzakaomarara maitiro masisitimu, uye iyo amplification kumhanya inogona kusvika 2Kb/min, iyo inonyanya kukodzera Direct PCR reaction.
Kit zvikamu
| Chikamu I | Buffer AL |
| Foregene Protease | |
| 6 × DNA Loading Buffer | |
| Chikamu II | 2 × PCR NyoreTMMusanganiswa(UNG) |
Zvimiro&zvakanakira
■Hapana nguva inopedza uye inodhura DNA kuchenesa
∎ Muenzaniso wekuda idiki, zvishoma se5mg mhuka yenyama inogona kuongororwa.
■ Hapana kurapwa kwakakosha sekukuya nekupwanya kunodiwa, uye kuvhiya kuri nyore.
■ Optimized PCR system inoita kuti PCR ive nemhando yepamusoro uye yakasimba kushivirira kune PCR reaction inhibitors.
■ Anti-kusvibiswa PCR system 2×PCR NyoreTMSanganisa (UNG) inobvisa zvinobudirira kusvibiswa kunokonzerwa nePCR zvigadzirwa uye inova nechokwadi cheiyo chaiyo uye nekurongeka kweamplification.
Kit parameters
Kushandisa: zvakasiyana-siyana zvemhuka.
DNA yakaburitswa kubva kumuenzaniso lysis: inoshandiswa chete sePCR template.
Iyo kit inogona kushandiswa kune zvinotevera zvinangwa: kuzivikanwa kwe transgenosis, mhuka genotyping, nezvimwe.
Kuyerera kwebasa
Storage uye Sherufu upenyu
Chikamu I chekiti ichi chinofanira kuchengetwa pa2-8℃.
Iyo reagent Buffer AL inogona kuchengetwa kwemwedzi gumi nemaviri pasi pemamiriro akaoma;inogona kuchengetwa pa2-8℃kuitira kuchengetedza kwenguva refu.
Reagent Foregene Protease ine yakasarudzika fomula.Kuti ive nechokwadi chekuita kwayo uye kugadzikana, ndapota chengeta pa4℃kwemwedzi gumi nemiviri.
Reagent 6×DNA Loading Buffer inogona kuchengetwa pa4℃kana -20℃kwenguva refu.
Chikamu II chekiti ichi chinofanira kuchengetwa pa -20℃.
Reagent 2×PCR NyoreTMMix(UNG), kana ikashandiswa kazhinji, inogona zvakare kuchengetwa pa4℃yekuchengetedza kwenguva pfupi (shandisa mukati memazuva gumi).
