Animal Tissue Direct PCR kit (isina Sampling Zvishandiso) Protocol
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 NyoreTMMusanganiswa une kushivirira kwakasimba kuPCR reaction inhibitors, uye inogona kushandisa lysate yemuenzaniso kuti iongororwe setemplate yeanoshanda uye chaiwo amplification.Iyi reagent ine Foregene D-Taq DNAPolymerase, dNTPs, MgCl.2, reaction buffer, PCR optimizer uye stabilizer.
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.
Zvinotsanangurwa
50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns
Kit zvikamu
Chikamu I | Buffer AL |
Foregene Protease | |
6 × DNA Loading Buffer | |
Chikamu II | 2 × PCR NyoreTMMix |
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.
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 NyoreTMMusanganiswa unogona kuchengetwa kwemwedzi gumi nemiviri pa -20℃;kana ikashandiswa kazhinji, inogona zvakare kuchengetwa pa4℃yekuchengetedza kwenguva pfupi (shandisa mukati memazuva gumi).