Hanyoyi na asali na Agarose Gel Electrophoresis (1)

1. Rarrabawa

Gel electrophoresis an kasu kashi a tsaye iri (ciki har da ginshiƙai gels da slab gels) da kuma kwance iri (yafi slab gels) (Hoto 6-18). Gabaɗaya, rarrabuwa a tsaye ya ɗan fi girma a kwance, amma shirye-shiryen gel a kwance yana da aƙalla fa'idodi guda huɗu: akwai tallafi a ƙarƙashin dukkan gel ɗin, yana ba da damar yin amfani da agarose mai ƙarancin hankali; yana yiwuwa a shirya faranti gel na agarose na ƙayyadaddun bayanai daban-daban; gel shirye-shiryen da samfurin loading sun fi dacewa; ɗakin electrophoresis yana da sauƙi don ginawa kuma yana da tsada. Tunda ana yin electrophoresis a kwance tare da farantin gel na agarose da aka nutsar da shi kusan 1mm ƙasa da farfajiyar buffer electrophoresis, ana kuma kiransa electrophoresis submerged.

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Electrophoresis Tank DYCP-31DN don agarose gel electrophoresis

2.Buffer System

A cikin rabuwar acid nucleic, yawancin tsarin suna ɗaukar tsarin ci gaba. Abubuwan buffer electrophoresis da aka fi amfani da su sun haɗa da TBE (0.08mol/L Tris·HCl, pH 8.5, 0.08mol/L boric acid, 0.0024mol/L EDTA) buffer da THE (0.04mol/L Tris·HCl, pH 7.8, 0.02mol/L sodium acetate, 0.0018mol/L EDTA) buffer. Ana shirya waɗannan buffer gabaɗaya azaman mafita na hannun jari na 10x kuma an diluted zuwa ƙimar da ake buƙata lokacin amfani. Matsakaicin ƙaura na madaidaiciya da DNA madauwari a cikin gel agarose sun bambanta tare da maƙerin da aka yi amfani da su. A cikin buffer, adadin ƙaura na DNA na layi ya fi na DNA madauwari, yayin da a cikin TBE buffer, akasin haka gaskiya ne.

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3.Shirin Agarose Gel

(1) Shiri Gel na Agarose na kwance

(a) Shirya abubuwan da ake buƙata na gel agarose ta amfani da buffer 1x electrophoresis.

(b) Zafafa agarose don kammala narkewa ko dai a cikin ruwan wanka mai tafasa, a kan injin maganadisu, ko a cikin microwave. Sanya maganin agarose zuwa 55 ° C kuma ƙara ethidium bromide (EB) rini zuwa matakin ƙarshe na 0.5 μg/ml.

(c) Rufe gefuna na gilashi ko faranti na acrylic tare da ƙaramin gel na agarose, ƙara tsefe, kuma sanya haƙoran tsefe kamar 0.5 ~ 1.0 mm sama da farantin.

(d) Zuba maganin gel na agarose da aka narke a cikin gilashin ko acrylic mold (kauri ya dogara da girman samfurin DNA), guje wa gabatarwar kumfa na iska. Bar shi ya ƙarfafa ta halitta a zafin jiki.

(e) Cire tsefe a tsanake bayan gama ƙarfi. Ƙara adadin da ya dace na buffer electrophoresis zuwa tankin gel, tabbatar da cewa gel farantin yana nutsewa game da 1 mm a ƙasa da saman buffer electrophoresis.

(2) Shiri Agarose Gel a tsaye

(a) Cire maiko ko saura daga faranti ta hanyar wankewa da ethanol.

(b) Sanya faranti na sarari tsakanin madatsun ruwa na gaba da na baya, daidaita gefuna na faranti tare da madatsun gaba da baya, sannan a tsare su da matsi.

(c) Ƙara agarose 2% a cikin madaidaicin 1x tsakanin gefuna na faranti don samar da filose mai tsayi cm 1 a ƙasan ɗakin simintin gel.

(d) Zuba gel ɗin agarose mai narkewa a cikin abin da ake so, wanda aka shirya a cikin buffer 1x, cikin ɗakin gel har zuwa 1 cm a ƙasan saman.

(e) Saka tsefe, guje wa tarko kumfa a ƙarƙashin haƙoran tsefe. Wani lokaci, wrinkles na iya bayyana a tsefe hakora a lokacin agarose gel sanyaya; a irin wannan yanayin, kawai a ƙara ɗan narke agarose a saman don ƙarfafa shi.

(f) Cire tsefe. Don hana zubewar buffer a cikin ramin lodi, rufe haɗin tsakanin farantin gel na agarose da ɗakin electrophoresis tare da 2% agarose kuma ƙara adadin da ake buƙata na buffer.

(g) Ƙara 1x electrophoresis buffer zuwa gel chamber.

(h) A hankali ɗora samfuran DNA a kan gel agarose da ke ƙarƙashin ma'ajin.

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Ƙarin bayani game da ainihin ilimin game da agarose gel electrophoresis, za mu raba mako mai zuwa. Yi fatan waɗannan bayanan su zama masu taimako ga gwajin ku.

Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) ya ƙware a masana'antar kayan aikin electrophoresis fiye da shekaru 50 tare da ƙungiyar ƙwararrun ƙwararrunmu da cibiyar R&D. Muna da amintaccen kuma cikakken layin samarwa daga ƙira zuwa dubawa, da ɗakunan ajiya, gami da tallafin talla. Babban samfuranmu sune Electrophoresis Cell (tanki / ɗakin), Samar da wutar lantarki na Electrophoresis, Mai watsawa na LED mai haske, Mai watsawa UV, Gel Image & Analysis System da dai sauransu.

Yanzu muna neman abokan tarayya, duka OEM electrophoresis tank da masu rarraba suna maraba.

Idan kuna da wani shirin siyan samfuranmu, da fatan a yi shakka a tuntuɓe mu. Kuna iya aiko mana da sako ta imel[email protected]ko[email protected], ko kuma a kira mu a +86 15810650221 ko ƙara Whatsapp +86 15810650221, ko Wechat: 15810650221.

Da fatan za a bincika lambar QR don ƙara akan Whatsapp ko WeChat.

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Lokacin aikawa: Dec-07-2023