2 × Taq PCR MasterMix Ⅱ

Rapid PCR premix tare da babban inganci da babban ƙarfin juriya.

2 × Taq PCR MasterMix Ⅱ shine sabon ingantaccen da haɓaka shirye-shiryen amfani da 2 × PCR tare da duk mahimman ɓangarori a cikin halayen PCR ban da samfurin DNA da firam ɗin.

Cat. A'a	Girman shiryawa
4993001	1ml ku
4993002	5x1ml ku
4992912	20x5x1 ku
4992913	5 × 1 ml
4992920	20 × 5 × 1ml
4992921	20 × 5 × 1ml

Aika mana imel

Bayanin samfur

Misalin Gwaji

Tambayoyi

Alamar samfur

Siffofin

Efficiency Ingantaccen haɓakawa: gutsutsuren DNA na masu girma dabam dabam (ƙasa da 5 kb) kuma ana iya haɓaka hanyoyin da kyau.
Sens Babban hankali: Kamar ƙasa da 10pg na gutsattsarin niyya za a iya haɓaka daga samfuran kwayoyin halitta.
Resistance Babban ƙarfin juriya: Don samfura tare da babban abun ciki na ƙazanta kamar samfuri mai tsattsauran ra'ayi/al'adun kwayan cuta, za a iya haɓaka guntun da ake nufi da sauƙi. Ayyukan polymerase ba zai shafar yawan daskarewa da narkewa ba.
Veni Mai dacewa don aikace -aikace: An shirya tsarin amsawa cikin sauƙi da sauri. Ƙarin ƙaramin ɓangaren yana ƙunshe da 3 ′ ƙarshen DA-overhang, wanda ya dace don cloning TA.

Musammantawa

Rubuta: Taken DNA polymerase
Samfurin: Samfurin da aka tsarkake/mai kauri/al'adun kwayan cuta
Samfura: > 10 pg
Girman guntu: <5kb ku
Aikace -aikace: Haɓaka PCR na gutsutsuren DNA, lakabin DNA, faɗaɗa na farko, ƙaddara jerin abubuwa, gano jigon manyan sikelin, gwajin PCR mai ƙima, gano DNA mai ganowa, da sauransu.

Duk samfuran ana iya keɓance su don ODM/OEM. Don cikakkun bayanai,don Allah danna Sabis na Musamman (ODM/OEM)

Na baya: 2 Mix GC mai arzikin PCR Mix

Na gaba: Tsarin PCR na Golden Easy (tare da fenti)

Hoto na 1. TIANGEN Taq MasterMix II da Taq Mix na yau da kullun daga Mai ba da TR ya haɓaka samfura daga maɓuɓɓuka daban -daban don gano ƙarfin juriya na reagents. Sakamakon ya nuna cewa samfuran TIANGEN na iya haɓaka gutsattsarin da aka nufa da su daga samfuran samfuran ƙwayoyin cuta da al'adun kwayan cuta, kuma juriya na danniya ya fi na Mai ba da TR. A: Samfurin danyen samfuri wanda TIANGEN TIANcombi DNA Lyse & Det PCR Kit ya fitar. Prp/DN: Hakar danyen da gano samfuran jinin ɗan adam. Shinkafa: hako danyen da gano samfuran shinkafa. B: PCR mai mulkin mallaka. Yankin PCR shine 700 bp.
M: TIANGEN Alamar III

Kyakkyawan duniya don samfura daga tushe daban -daban kuma tare da tsayin tsayi daban -daban
Hoto 2. An haɓaka gutsuttsarin hanyoyin daban da tsayi daban -daban ta amfani da TIANGEN Taq MasterMix II (A) da talakawa Taq Haɗin Mai ba da TK (B), Mai ba da kaya TR (C), Mai ba da V (D) da Mai ba da G (E) bi da bi. Sakamakon ya nuna cewa cikakkiyar aikin samfuran TIANGEN shine mafi kyau dangane da ƙarfin faɗaɗawa, keɓancewa da duniya baki ɗaya.M: TIANGEN Alamar III1: Soybean genomic DNA template (120 bp);

2-3: Samfurin kwayar halittar DNA na shinkafa (694 bp, 2258 bp);

4: Samfurin DNA na auduga genomic (200 bp);

5: Escherichia coli samfurin DNA na genomic (2298 bp);

6-7: Tsarin DNA na Mouse (1 kb, 2 kb);

8-10: Samfurin DNA na bera (1 kb, 2 kb, 2080 bp);

11-18: Samfurin DNA na ɗan adam (300 bp, 448 bp (GC%: 74.8%), 1100 bp, 750 bp,

1000 bp, 1090 bp (GC%: 70.4%), 2 kb, 4 kb)

Babban hankali
Hoto na 3. An ƙara girman ɗimbin bera da gutsutsuren DNA na ɗan adam ta amfani da TIANGEN Taq MasterMix II (A), talakawa Taq Haɗin Mai Bayarwa V (B) da Mai ba da TK (C), bi da bi, don gano ƙarfin haɓakawa. Sakamakon ya nuna cewa samfurin TIANGEN na iya haɓaka gutsattsarin abin da aka nufa daga samfuran ƙirar ƙasa har zuwa 0.01 ng, kuma hankalin sa ya fi na samfuran daga Mai Bayarwa V da TK.M: TIANGEN Alamar III, N: NTCTemplate shigar 1-8 : 200 ng, 100 ng, 50 ng, 20 ng, 10 ng, 1 ng, 0.1 ng, 0.01 ng.

Tambaya: Babu makaɗan ƙarawa

Samfurin A-1

Template Samfurin yana ɗauke da ƙazamar furotin ko masu hana Taq, da dai sauransu ——- Tsarkake samfurin DNA, cire ƙazantar furotin ko cire samfuri na DNA tare da kayan tsarkakewa.

■ Ƙaddamar da samfuri bai cika ba ——Ya dace a ƙara yawan zazzabin denaturation da tsawaita lokacin denaturation.

Deg ƙasƙantar da samfuri ——Ka sake shirya samfuri.

Bayanan Bayani na A-2

Quality Kyakkyawan ingancin fitila — —Re-synthesize primer.

Deg ƙasƙantar da kai na farko — —Aliquot babban firam ɗin taro zuwa ƙaramin ƙara don adanawa. Guji daskarewa da narkewa da yawa ko adanawa na 4 ° C na dogon lokaci.

Design Tsararrun ƙirar firamari (misali tsawon firamare bai isa ba, dimer da aka kafa tsakanin firam ɗin, da dai sauransu) -Rirar ƙira (ku guji samuwar dimer primer da tsarin sakandare)

A-3 MG²⁺maida hankali

■ Mg²⁺ maida hankali yayi ƙasa sosai ——Yin haɓaka Mg daidai²⁺maida hankali: Inganta Mg²⁺ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg²⁺ maida hankali ga kowane samfuri da share fage.

A-4 Ƙara yawan zafin jiki

Zazzabi mai ƙima yana shafar ɗaurin firamare da samfuri. —— Rage zafin zafin zafin jiki da inganta yanayin tare da ɗimbin digiri na 2 ° C.

A-5 Tsawaita lokacin

■ Gajerun lokacin ƙara —— Ƙara lokacin ƙarawa.

Tambaya: Tabbatacce

Phenomena: Samfuran marasa kyau suma suna nuna makasudin jerin makasudin.

A-1 Gurɓatar PCR

Tsallake gurɓataccen jerin abubuwan da aka yi niyya ko samfuran ƙarfafawa ——Ayi hankali kada a ɗora samfurin da ke ɗauke da jerin manufa a cikin samfurin mara kyau ko zubar da su daga bututun ƙarfe. Yakamata reagents ko kayan aiki suyi aiki da kan su don kawar da acid nucleic da ke akwai, kuma yakamata a ƙaddara wanzuwar gurɓatarwa ta hanyar gwajin sarrafawa mara kyau.

Contamin Gurɓatawa mai gurɓatawa ——Aliquot reagents da adanawa a ƙaramin zafin jiki.

A-2 Firayim Ministar

■ Mg²⁺ maida hankali yayi ƙasa sosai ——Yin haɓaka Mg daidai²⁺ maida hankali: Inganta Mg²⁺ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg²⁺ maida hankali ga kowane samfuri da share fage.

Tsarin ƙirar da bai dace ba, kuma jerin maƙasudin yana da homology tare da jerin marasa manufa. ——Re-design primers.

Tambaya: Ƙarfafawa ba takamaimai ba

Phenomena: Ƙungiyoyin haɓaka PCR ba su dace da girman da ake tsammani ba, ko babba ko ƙarami, ko kuma wani lokacin duka takamaiman ƙungiyoyin haɓakawa da ƙamus ɗin ba na musamman ba.

Farashin A-1

Ƙididdiga ta musamman

——Re-design primer.

Concentration Mahimmancin taro ya yi yawa sosai ——Ya ƙara haɓaka zafin denaturation da tsawaita lokacin denaturation.

A-2 MG²⁺ maida hankali

M Mg²⁺ maida hankali ya yi yawa sosai ——Ya dace a rage maida hankali Mg2+: Inganta Mg²⁺ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg²⁺ maida hankali ga kowane samfuri da share fage.

A-3 Polymerase mai ƙarfi

Yawan adadin enzyme mai yawa ——Rage adadin enzyme yadda yakamata a tsakanin 0.5 U.

A-4 Ƙara yawan zafin jiki

Temperature Zazzabin ƙoshin ya yi ƙasa da yawa ——Ya dace a ƙara yawan zafin zafin ɗin ko a yi amfani da hanyar ƙara matakin biyu

A-5 PCR hawan keke

Cy Yawan hawan PCR da yawa —— Rage yawan hawan PCR.

Tambaya: Ƙungiyoyi masu ƙyalli ko shafa

Farashin A-1—— Bayanai marasa kyau ——Rayyana ƙirar fitila, canza matsayi da tsawon fitilar don haɓaka takamaiman sa; ko yin PCR da aka ƙera.

A-2 Template DNA

—— Samfurin ba shi da tsarki ——Ka tsarkake samfur ko cire DNA tare da kayan tsarkakewa.

A-3 MG²⁺ maida hankali

——Mg²⁺ maida hankali yayi yawa ——Ya rage Mg daidai²⁺ maida hankali: Inganta Mg²⁺ maida hankali ta hanyar jerin halayen daga 1 mM zuwa 3 mM tare da tazara na 0.5 mM don ƙayyade mafi kyawun Mg²⁺ maida hankali ga kowane samfuri da share fage.

A-4 dNTP

——Tinganin dNTPs sun yi yawa ——Rage taro na dNTP yadda yakamata

A-5 Ƙara yawan zafin jiki

—— Ƙaramin zafin zafin ƙanƙara mai zafi ——Ya dace a ƙara yawan zafin zafin

Hanyoyin A-6

——Yawan hawan keke da yawa ——Ya inganta lambar zagayowar

Tambaya: Nawa samfuri nawa ne ya kamata a ƙara a cikin tsarin amsawa na 50 μl PCR?

Tambaya: Yadda za a fadada dogayen gutsutsuren?

Mataki na farko shine zaɓi polymerase da ya dace. Taq polymerase na yau da kullun ba zai iya sake karantawa ba saboda rashin aikin fitowar 3'-5 ', kuma rashin daidaituwa zai rage ƙimar fadada guntu. Sabili da haka, polymerase na Taq na yau da kullun ba zai iya haɓaka gutsattsarin da ya fi girma fiye da 5 kb ba. Taq polymerase tare da gyare -gyare na musamman ko wani babban aminci polymerase yakamata a zaɓi don haɓaka ingantaccen haɓakawa da saduwa da buƙatun ƙaramin guntu. Bugu da ƙari, ƙaramin guntun gutsutsuren kuma yana buƙatar daidaita daidaiton ƙirar fitila, lokacin denaturation, lokacin faɗaɗawa, pH mai ɓoyewa, da sauransu. Don hana lalacewar samfuri, lokacin denaturation a 94 ° C yakamata a rage zuwa 30 sec ko ƙasa da kowane zagayowar, kuma lokacin haɓaka zafin jiki zuwa 94 ° C kafin haɓaka ya zama ƙasa da 1 min. Haka kuma, saita zafin zafin a kusan 68 ° C da kuma tsara lokacin tsawaita gwargwadon ƙimar 1 kb/min na iya tabbatar da ingantaccen ingantaccen guntun gutsutsuren.

Tambaya: Yadda za a inganta amincin amincin PCR?

Ana iya rage ƙimar kuskuren haɓaka PCR ta amfani da polymerases DNA daban -daban tare da babban aminci. Daga cikin duk polymerases na Taq DNA da aka samo zuwa yanzu, enfuz enzyme yana da mafi ƙarancin ƙimar kuskure da mafi aminci (duba tebur da aka makala). Baya ga zaɓin enzyme, masu bincike na iya ƙara rage ƙimar maye gurbi ta PCR ta hanyar inganta yanayin amsawa, gami da haɓaka abun da ke ciki, maida hankali na polymerase mai ɗorewa da haɓaka lambar sake zagayowar PCR.

Rubuta saƙonka a nan ka aika mana