Mouse Genotyping Kit

Cat No: HCR2021A

Wannan samfurin kit ne da aka ƙera don saurin gano genotypes na linzamin kwamfuta, gami da hakar ɗanyen DNA da tsarin haɓaka PCR.Ana iya amfani da samfurin don haɓaka PCR kai tsaye daga wutsiyar linzamin kwamfuta, kunne, yatsan yatsa da sauran kyallen takarda bayan sauƙaƙe ta hanyar Lysis Buffer da Proteinase k.Babu narkewa na dare, cirewar phenol-chloroform ko tsarkakewar shafi, wanda ke da sauƙi kuma yana rage ɗaukar lokaci na gwaji.Samfurin ya dace da haɓaka ɓangarorin da aka yi niyya har zuwa 2kb da halayen PCR masu yawa tare da har zuwa nau'i-nau'i na 3.2 × Mouse Tissue Direct PCR Mix yana ƙunshe da DNA polymerase wanda aka kirkira ta asali, Mg²⁺, dNTPs da ingantaccen tsarin buffer don samar da ingantaccen haɓaka haɓakawa da haƙuri mai hanawa, ta yadda za'a iya aiwatar da halayen PCR ta ƙara samfuri da masu haɓakawa da sake sabunta samfurin zuwa 1 ×.Samfurin PCR da aka haɓaka tare da wannan samfur yana da sanannen tushe “A” a ƙarshen 3′ kuma ana iya amfani dashi kai tsaye don cloning TA bayan tsarkakewa.

Abubuwan da aka gyara

Yanayin Ajiya

Ya kamata a adana samfuran a -25 ~ -15 ℃ don shekaru 2.Bayan narke, Lysis Buffer za a iya adana a 2 ~ 8 ℃ don gajeren lokaci mahara amfani, da kuma Mix da kyau lokacin amfani.

Aikace-aikace

Wannan samfurin ya dace da binciken ƙwanƙwasa linzamin kwamfuta, gano transgenic, genotyping da sauransu.

Siffofin

1.Aiki mai sauƙi: babu buƙatar cire DNA na genomic;

2.Wide aikace-aikace: dace da kai tsaye ƙarawa daban-daban linzamin kwamfuta kyallen takarda.

Umarni

1.Sakin DNA na genomic

1) Shiri na lysate

An shirya lysate nama bisa ga adadin samfuran linzamin kwamfuta da za a lysed (ya kamata a shirya lysate nama akan rukunin yanar gizon gwargwadon sashi kuma a haɗe shi sosai don amfani), kuma adadin reagents da ake buƙata don samfurin guda ɗaya shine kamar haka:

2) Samfurin Shiri da Lysis

An Shawarar Amfani da Nama

Ɗauki samfurin nama na linzamin kwamfuta da ya dace a cikin bututun centrifuge mai tsabta, ƙara 200μL na sabbin ƙwayoyin nama zuwa kowane bututun centrifuge, vortex da girgiza, sannan a sanya shi a 55 ℃ na 30mins, sannan zafi a 98 ℃ na 3mins.

3) Tsare-tsare

Ki girgiza lysate da kyau kuma a yi ta a 12,000 rpm na 5mins.Za a iya amfani da maɗaukakin abu azaman samfuri don haɓaka PCR.Idan ana buƙatar samfuri don ajiya, canja wurin supernatant zuwa wani bututu mai bakararre kuma adana a -20 ℃ na makonni 2.

2.PCR Amplification

Cire 2 × Mouse Tissue Direct PCR Mix daga -20 ℃ kuma narke akan kankara, haɗuwa da juye kuma shirya tsarin amsawar PCR bisa ga tebur mai zuwa (aiki akan kankara):

Lura:

a) Adadin da aka ƙara bai kamata ya wuce 1/10 na tsarin ba, kuma idan an ƙara da yawa, ana iya hana haɓakar PCR.

Sharuɗɗan PCR da aka Shawarar

Lura:

a) Annealing zafin jiki: Dangane da darajar Tm na firamare, ana ba da shawarar saita zafin jiki na ƙarami zuwa ƙaramin ƙimar Tm na firam + 3 ~ 5 ℃.

Matsalolin gama gari da Magani

1.Babu tube da aka yi niyya

1) Yawan lysis samfurin.Zaɓi mafi dacewa adadin samfuri, yawanci bai wuce 1/10 na tsarin ba;

2) Girman samfurin da yawa.Tsarma da lysate sau 10 sannan kuma ƙarawa, ko rage girman samfurin kuma sake sakewa;

3) Samfurin nama ba sabo ba ne.Ana ba da shawarar yin amfani da samfuran nama sabo;

4) Rashin inganci mai inganci.Yi amfani da DNA na genomic don haɓakawa don tabbatar da ingancin firamare da haɓaka ƙirar firamare.

2.Ƙwaƙwalwar da ba takamaiman ba

1) Zazzaɓin zafi ya yi ƙasa da ƙasa kuma lambar sake zagayowar ta yi yawa.Ƙara yawan zafin jiki na annealing kuma rage yawan hawan keke;

2) Matsalolin samfuri ya yi yawa.Rage adadin samfuri ko tsarma samfurin sau 10 bayan haɓakawa;

3) Rashin ƙayyadaddun ƙayyadaddun ƙa'idodi.Inganta ƙirar farko.

Bayanan kula

1.Don kauce wa ƙetare gurɓata tsakanin samfurori, ya kamata a shirya kayan aikin samfuri da yawa, kuma ana iya tsaftace saman kayan aikin tare da 2% sodium hypochlorite bayani ko nucleic acid cleaner bayan kowane samfurin idan ana buƙatar maimaita amfani.

2.Ana ba da shawarar yin amfani da sabbin kyallen linzamin kwamfuta, kuma girman samfurin bai kamata ya yi girma da yawa ba don guje wa shafar sakamakon haɓakawa.

3.Lysis Buffer ya kamata ya guje wa daskare-narke akai-akai, kuma ana iya adana shi a 2 ~ 8 ℃ don amfani da yawa na ɗan gajeren lokaci.Idan an adana shi a ƙananan zafin jiki, hazo na iya faruwa, kuma dole ne a narkar da shi sosai kafin amfani.

4.Ya kamata PCR Mix ya guje wa daskare-narke akai-akai, kuma ana iya adana shi a 4 ℃ don maimaita amfani na ɗan lokaci.

5.Wannan samfurin don bincike ne na gwaji na kimiyya kawai kuma bai kamata a yi amfani da shi ba wajen ganewar asibiti ko magani.