Robustart Taq DNA Polymerase

Robustart Taq DNA Polymerase shine farkon DNA polymerase mai zafi.Wannan samfurin ba zai iya kawai mafi kyawun hana abin da ba takamaiman abin da ya haifar ba ta hanyar rashin takamaiman takamammen abubuwan da aka haɗa na al'ada ko tari a cikin tsarin shirye-shiryen PCR da haɓakawa.Sabili da haka, yana da ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun bayanai kuma ya fi tasiri don haɓaka ƙarancin ƙima, kuma ya dace da haɓaka haɓakawar haɓakawa na PCR.Bugu da ƙari, wannan samfurin yana da kyakkyawan aiki, kuma ana iya samun sakamako mai ƙarfi a cikin nau'ikan halayen PCR daban-daban.

Abubuwan da aka gyara

1.5 U/μL Robustart Taq DNA polymerase

2.10 × PCR Buffer II (Mg²+ kyauta) (na zaɓi)

3.25 mMgCl₂(na zaɓi)

* 10 × PCR Buffer II (Mg²+ kyauta) bashi da dNTP da Mg²+, da fatan za a ƙara dNTPs da MgCl₂lokacin shirya tsarin amsawa.

Abubuwan da aka Shawarar

1.Saurin haɓakawa.

2.Ƙarawa da yawa.

3.Kai tsaye haɓaka jini, swabs, da sauran samfurori.

4.Gano cututtuka na numfashi.

Yanayin Ajiya

-20 ° C don ajiya na dogon lokaci, ya kamata a gauraye da kyau kafin amfani, kauce wa daskare-narke akai-akai.

*Idan ruwan sama ya faru bayan sanyaya, al'ada ce;ana bada shawara don daidaita zuwa zafin jiki kafin haɗuwa da amfani.

Ma'anar Naúrar

Ɗaya daga cikin naúrar aiki (U) an bayyana shi azaman adadin enzyme wanda ya haɗa 10 nmol na deoxyribonucleotide a cikin kayan acid-insoluble a 74 ° C na 30mins ta amfani da DNA na maniyyi mai kunnawa azaman samfuri/primer.

Kula da inganci

1.SDS-PAGE tsarki na electrophoretic fiye da 98%.

2.Amplification hankali, sarrafa tsari-zuwa-tsari, kwanciyar hankali.

3.Babu wani aiki na nuclease, babu exogenous endonuclease ko exonuclease gurbatawa

Umarni

Saitin Amsa

Bayanan kula:

1) a.Makullin baya ƙunshi dNTP da Mg²+, da fatan za a ƙara dNTPs da MgCl₂lokacin shirya tsarin amsawa.

2) b.Idan aka yi amfani da shi don qPCR/qRT-PCR, ya kamata a ƙara ƙwaƙƙwaran bincike zuwa tsarin amsawa.Yawancin lokaci, ƙaddamarwa na ƙarshe na 0.2 μM zai ba da sakamako mai kyau;idan aikin amsawa ba shi da kyau, ana iya daidaita maida hankali na farko a cikin kewayon 0.2-1 μM.Yawan binciken binciken ana inganta shi a cikin kewayon 0.1-0.3 μM.Za a iya yin gwaje-gwajen hankali don nemo mafi kyawun haɗe-haɗe na firamare da bincike.

Ƙa'idar hawan keke na thermal

Bayanan kula

1.Adadin haɓakawa na DNA polymerase mai sauri kada ya zama ƙasa da 1 kb/10 s.Hawan zafin jiki da ƙimar faɗuwa, yanayin kula da zafin jiki da ingancin tafiyar da zafi na kayan aikin PCR daban-daban sun bambanta sosai, don haka ana ba da shawarar haɓaka ingantattun yanayin amsawa don takamaiman kayan aikin PCR mai sauri.

2.Tsarin yana da sauƙin daidaitawa, tare da ƙayyadaddun ƙayyadaddun bayanai da ƙwarewa.

3.Ya dace da amfani azaman babban ji na PCR gano reagents, kuma ana iya amfani dashi a cikin halayen haɓakawa na PCR mai yawa.

4.5′→3′ Ayyukan polymerase, 5′→3′ Ayyukan exonuclease;babu 3′→5′ aikin exonuclease;babu aikin tantancewa.

5.Ya dace da gwaji da ƙididdigewa na PCR da RT-PCR.

6.Ƙarshen 3′ na samfurin PCR shine A, wanda za'a iya haɗa shi kai tsaye zuwa cikin vector T.

7.Ana ba da shawarar hanyar matakai uku don masu farawa tare da ƙananan yanayin zafi ko don haɓaka gutsuttsura fiye da 200 bp.