Robustart Taq DNA Polymerase

Inombolo yekati:HC1014A

Iphakheji: 0.1ml/1ml/5ml

Robustart Taq DNA Polymerase sisiqalo eshushu DNA polymerase.

Thumela uMbuzo

Ingcaciso yeMveliso

Iinkcukacha zemveliso

Robustart Taq DNA Polymerase sisiqalo eshushu DNA polymerase.Le mveliso ayinakunqanda ngcono kuphela ukusabela okungachanekanga okubangelwa kukufakwa okungangqalanga kwe-primers okanye ukuhlanganiswa kwe-primer kwinkqubo yokulungiswa kwenkqubo ye-PCR kunye nokukhulisa.Ke ngoko, inobuchwephesha obubalaseleyo kwaye iyasebenza ngakumbi ekwandiseni itemplates zoxinaniso oluphantsi, kwaye ifanelekile kwi-multiplexed PCR yokusabela kokukhulisa.Ngapha koko, le mveliso inokusebenziseka okuhle kakhulu, kwaye iziphumo zokukhulisa ezizinzileyo zinokufumaneka kwiindidi ezahlukeneyo zokusabela kwePCR.

Ngaphambili: Superstart Taq DNA Polymerase HC1013A

Okulandelayo: I-M-MLV Reverse Transcriptase HC2003B

Amacandelo

1.5 U/μL Robustart Taq DNA polymerase

2.10 × PCR Buffer II (Mg²+ simahla) (ukhetho)

3.25 mM MgCl₂(ukhetho)

* 10 × PCR Buffer II (Mg²+ simahla) ayiqulathanga dNTP kunye neMg²+, nceda wongeze idNTPs kunye neMgCl₂xa ulungiselela inkqubo yokusabela.

Izicelo ezicetyiswayo

1.Ukukhulisa ngokukhawuleza.

2.Ukwandisa okuninzi.

3.Ukwandiswa ngokuthe ngqo kwegazi, i-swabs, kunye nezinye iisampuli.

4.Ukufunyanwa kwezifo zokuphefumla.

Imeko yoGcino

-20°C kwindawo yokugcina ixesha elide, kufuneka ixutywe kakuhle phambi kokusetyenziswa, kunqande ukunyibilika komkhenkce rhoqo.

*Ukuba imvula iyenzeka emva kwefriji, kuqhelekile;kucetyiswa ukulinganisa ubushushu begumbi ngaphambi kokuxuba kunye nokusetyenziswa.

Inkcazelo yeyunithi

Iyunithi enye esebenzayo (U) ichazwa njengobungakanani be-enzyme edibanisa i-10 nmol ye-deoxyribonucleotide kwizinto ezingenayo i-asidi kwi-74 ° C ye-30mins isebenzisa i-salmon yesidoda se-DNA esebenzayo njenge-template / primer.

Ulawulo lwemeko

1.I-SDS-PAGE ukucoceka kwe-electrophoretic enkulu kune-98%.

2.Ukwandisa ubuntununtunu, i-batch-to-batch control, uzinzo.

3.Akukho msebenzi we-nuclease exogenous, akukho endonuclease exogenous okanye exonuclease ungcoliseko

Imiyalelo

Ukuseta iRection

Amacandelo	Umthamo (μL)	Ukugxininiswa kokugqibela
10 × PCR Buffer II (Mg²+ simahla)^a	5	1×
dNTPs (10mM idNTP nganye)	1	200 μM
25 mM MgCl₂	2-8	1-4 mm
I-Robustart Taq DNA Polymerase (5U/μL)	0.25-0.5	1.25-2.5 U
I-25 × i-Primer mix^b	2	1×
Isifanekiso	-	＜ 1 μg/impendulo
ddH₂O	Ukuya kwi50	-

Amanqaku:

1) a.Isithinteli asiqulathanga idNTP kunye neMg²+, nceda wongeze idNTPs kunye neMgCl₂xa ulungiselela inkqubo yokusabela.

2) b.Ukuba isetyenziselwa i-qPCR/qRT-PCR, iiprobe zefluorescent kufuneka zongezwe kwindlela yokusabela.Ngokuqhelekileyo, i-primer concentration yokugqibela ye-0.2 μM iya kunika iziphumo ezilungileyo;ukuba i-reaction performance ayilunganga, i-primer concentration ingalungiswa kuluhlu lwe-0.2-1 μM.I-probe concentration idla ngokulungiswa kuluhlu lwe-0.1-0.3 μM.Imifuniselo yegradient yoxinaniso inokwenziwa ukufumana eyona ndibaniselwano yeprimer kunye neprobe.

Iprothokholi yebhayisekile eshushu

I-PCR rhoqoinkqubo
Inyathelo	Ubushushu	Ixesha	Imijikelo
I-denaturation yangaphambili	95℃	1-5 imiz	1
I-Denaturation	95℃	10-20 imizuzwana	40-50
Ukwandiswa / Ukwandiswa	56-64℃	20-60 umzuzwana	40-50

I-PCR ekhawulezayoinkqubo
Inyathelo	Ubushushu	Ixesha	Imijikelo
I-denaturation yangaphambili	95℃	30 imizuzwana	1
I-Denaturation	95℃	1-5 imizuzwana	40-45
Ukwandiswa / Ukwandiswa	56-64℃	5-20 imizuzwana	40-45

Amanqaku

1.Izinga lokukhulisa i-polymerase ye-DNA ekhawulezayo akufunekanga ibe ngaphantsi kwe-1 kb/10 s.Ukunyuka kobushushu kunye nesantya sokuwa, imo yokulawula ubushushu kunye nokusebenza kakuhle kobushushu bezixhobo ezahlukeneyo zePCR kuyahluka kakhulu, ngoko kuyacetyiswa ukuba kunyuswe iimeko zokusabela ezifanelekileyo kwisixhobo esithile esikhawulezayo sePCR.

2.Inkqubo iguquguquka kakhulu, kunye neenkcukacha eziphezulu kunye novakalelo.

3.Ilungele ukusetyenziswa njengobuntununtunu obuphezulu bokufunyanwa kwe-PCR, kwaye ingasetyenziswa kwi-multiplex ye-PCR yokusabela kokukhulisa.

4.5′→3′ umsebenzi wepolymerase, 5′→3′ umsebenzi we-exonuclease;hayi 3′→5′ exonuclease umsebenzi;akukho msebenzi wokuvavanya.

5.Ifanelekile kuvavanyo lomgangatho kunye nobungakanani be-PCR kunye ne-RT-PCR.

6.Ukuphela kwe-3 yemveliso ye-PCR ngu-A, enokuthi ifakwe ngokuthe ngqo kwi-T vector.

7.Indlela yamanyathelo amathathu iyacetyiswa kwii-primers ezinamaqondo aphantsi okutshisa i-annealing okanye ukukhulisa amaqhekeza angaphezulu kwe-200 bp.