Multiplex Mataki Daya RT-qPCR Premix-UNG
Saukewa: HCB5145A
Multiplex One Mataki na RT-qPCR Probe Kit (UDG Plus) kit ɗin PCR ne mai yawa bisa RNA azaman samfuri.A cikin aiwatar da gwajin, an yi jujjuya rubutu da PCR mai ƙididdigewa a cikin bututu guda ɗaya, wanda ya sauƙaƙa aikin gwaji kuma ya rage haɗarin kamuwa da cuta.A cikin wannan kit ɗin, cDNA na farko an haɗa shi da kyau ta hanyar Reverse Transcriptase mai jure zafin zafi kuma HotStart Tag DNA Polymerase ya haɓaka da yawa.Kit ɗin yana ƙunshe da ingantaccen buffer MP, haɗin enzymes, da sauransu. Maganin buffer ya riga ya ƙunshi Mg2+da dNTP.Bugu da ƙari, abubuwan da za su iya hana haɓakar haɓakar PCR marasa ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun haɓakawa da haɓaka haɓaka haɓakar halayen qPCR da yawa ana ƙara, waɗanda zasu iya tabbatar da ingancin haɓakawa da aiwatar da haɓaka haɓakawa da yawa.An ƙara tsarin dUTP/UDG don hana haɗarin gurɓataccen iska.
Abubuwan da aka gyara
1. Buffer
2. Enzyme Mix
Ƙayyadaddun bayanai
| Zafafan Farawa | Gina-in zafi farawa |
| Hanyar ganowa | Gano na farko-bincike |
| Hanyar PCR | Mataki ɗaya RT-qPCR |
| Polymerase | Taq DNA polymerase |
| Nau'in samfurin | DNA |
Yanayin Ajiya
Ana jigilar samfurin tare da busassun ƙanƙara kuma ana iya adana shi a -25 ~ -15 ℃ na shekara 1.Ya kamata a guji akai-akaidaskare-narke.Ana ba da shawarar yin ajiya daban.
Umarni
1.Martani Tsari
| Abubuwan da aka gyara | girma (μL) | Ƙarshe Tattaunawa |
| 2 × MP Buffer | 12.5 | 1× |
| Enzyme Mix | 1 | - |
| Haɗin Farko/Bincike (2.5 μM) | 3 | 0.3 m |
| Samfura RNA | 1-10 | - |
| RNase H2O Kyauta | zuwa 25 | - |
Bayanan kula:
Tabbatar ku haɗu da kyau kafin amfani, kauce wa wuce kima kumfa wanda tashin hankali ya haifar.
a.Mahimmanci na farko: Haɗawar farko gami da maɗaukakiyar share fage, ya danganta da yanayin mafi kyawun maida hankali mai ƙila tsakanin 0.l da 1.0μM.
b.Matsakaicin bincike: Haɗin bincike gami da bambancin alamar bincike na multix, ya danganta da yanayin mafi kyawun tattarawar binciken ƙila tsakanin 0.05 da 0.5μM.
c.Tsarin samfuri: qPCR yana da matukar kulawa kuma ana ba da shawarar a tsoma samfurin.Ƙimar Ct mai sarrafawa ta dace tsakanin 20 zuwa 35.
d.Shirye-shiryen tsarin: Da fatan za a shirya a cikin teburin aiki mai tsabta mai tsabta, da pipettor da bututu mai amsawa ba tare da ragowar nuclease ba;ana ba da shawarar yin amfani da shugaban bindiga tare da abubuwan tacewa.Guji gurbacewar giciye da gurɓataccen iska.
2. Ingantattun KekunaYarjejeniya
| Zagayowar mataki | Temp. | Lokaci | Zagaye |
| Juya rubutun | 50 ℃a | 20 min | 1 |
| Na farko-denaturation | 95 ℃ | 5 min | 1 |
| Amplification dauki | 95 ℃ | dakika 15 |
40-45 |
| 60 ℃ b | 30 seconds c |
Bayanan kula:
a.Juya fassarar: Zazzabi na iya zaɓar 42°C ko 50°C.
b.Amplification Amplification: Ana daidaita zafin jiki bisa ga ƙimar Tm na abubuwan da aka tsara.
c.Samun siginar Fluorescence: Da fatan za a saita tsarin gwaji bisa ga buƙatun littafin kayan aiki.
Bayanan kula
Da fatan za a saka PPE da ake buƙata, kamar sut ɗin lab da safar hannu, don tabbatar da lafiyar ku da amincin ku.
