Amanyathelo okhuseleko lwe-Polyacybuster gel Electrophoresis

· i-polyacybuslamIjeli kufuneka i-acrylamer monomer, i-polymerizazation iqala izinto, i-catalyst, kunye nelungelo letyuwa kunye nomxube we-buffer kunye.

· i-acrysidekunye ne-bis (n, n '- i-methylene i-acrymide) yi-monomer ye-molmer ye-Gel Matrix.

I-AmAMMMoind Plasfaind iqale inkqubo yokubulala i-polymeriting. Ukuqulunqwa kweglu kufuna isisombululo se-10% ye-Ammonium sePromoum ​​senzelwa emanzini. Uninzi lolwazi lubonise isidingo sokusetyenziswa okusebenzayo. Nangona kunjalo, isisombululo se-10% sinokubekwa kwi-4 ℃ kwiiveki ezininzi ngaphandle kokuphulukana nomsebenzi. Yenza ukuya kwi-10 ml kwaye ulahle xa iglusi iyasilela ukuhlanganisa.

Ingcebiso: Ipesenti yei-acrysideKwindlela yokulandelelana kweglue kunye neprotheyini iglue ayifani. Ukuba usebenzisa i-Acrysam Acryside: Isisombululo se-BIS, qinisekisa ukuba ibhotile elungileyo.

I-N, N, N, N, n '- iTetramethyl Eslediamine) yibhotile ye-brown, ibekwe kwifriji. Yongeza ngaphambi kokuba utyelele iglu.

I-elekkyfophorander e-Incraphoressis isebenzise iglasi kwi-elektrophoressis kufuneka ihlanjwe ngaphambi nasemva kwexesha ngalinye. Emva kwe-elektrophoreris, hlamba ngebrashi ethambileyo kunye nelaphu ngamanzi ashushu asekweleyo, hlamba ngamanzi anezinzi kwaye ume ukuze wome.

Ukuqina nothuli kunokubangela i-polymer ye-holymer. Phambi kwe-elektrophoris, coca iplate yeglasi kunye neglasi esicocekileyo kwaye uyisule ngebrashi ethambileyo. Hlamba ngamanzi anqatyisiweyo kwaye womile ngokupheleleyo ngephepha lokusula iphepha. Ukuhamba nge-70% ye-ethanol ngaphambi kokuba ususe iphepha kunceda ukucoca kunye nokukhawulezisa ukomisa. Yongeza iisampulu ze-abrymide ngokulandelelana: I-BIS, amanzi, isisombululo se-buffer, i-Ammonium PersiperFete, igcwele. Shake kakuhle kwaye ugalele kwangoko.

Akuyimfuneko kwi-Pardos Polsacryside ngaphambi kokugcwaliswa kwePolymeri. .

I-GLUE YOKUGQIBELA IIKHONO ZEEMPAWU.

Iphepha elimnyama lephepha elimnyama kwibhokisi engezantsi, imvelaphi emnyama ukwenza ukuba i-Hole yesampulu ukubona ngokucacileyo.
I-Buffer egcwalise i-buffer, ngaphezulu kwe-colloid.
Ukukhanya kukukhanya, jika izibane, makukhanyise ukukhanya kwe-colloid. Zoba isampulu emphelweni.
· · · Pelisa isixhobo sokuzipho.
I-Mu 1-200 MU 1 indawo yokuhamba inokusetyenziswa kumanqaku amaninzi kwisampulu. Kwimingxunya enesampulu encinci kakhulu (ngaphantsi kwe-5μ), intloko ende ye-pipette esetyenziselwa ukulandelelana kweglue ilunge ngakumbi.

Ukutshintsha nje nje kwisampulu, ukuphefumla kancinci kancinci. Isampulu inokubonakala inamathela nge-glycerin, kwaye impompo ngokukhawuleza inokuzoba iibhubhu ze-moya kwintloko yombhobho.

Iisakhelo emva kokuphefumla intloko yokuphefumla, iya kushukumisa i-fluid intloko ngobunono kumda wombhobho okanye iphepha le-suck lincancisa i-scolets ngaphandle kwamaconsi. Lumka ungancancisa isampulu.

Beka isampulu kwimodeli yesampulu

Isixhobo sokutshintsha isixhobo ukugcina uxinzelelo elincinci, yenza iisampulu zihamba kancinci intloko yolwelo.
Intloko ye-Pipet ifakwe kwi-buffer, ingaphezulu kancinci kunemingxunya yendawo, gcina uxinzelelo olufanelekileyo. Incam yomphunga inokufakwa kumngxunya omncinci.
· · Meely kwaye ngokuthe ngcembe iisampuli ngaphandle. I-PIPTE Ingcebiso ibekwe ngaphezulu kwenqaku lesampulu, kwaye isampulu iya kutsiba emngxunyeni. Vumela isampulu inyibilikisi ukugcwalisa umngxunya wesampulu endaweni yokutyhala.
Ukwehla kokugqibela kwesampulu ngentloko yolwelo, ulwelo luya kufikelela kumlenze wesibini, ngokuthe ngcembe luphakamisa ukuhamba ulwelo, ukuphuma kwi-buffer

Ungayenza njani i-GLUU YOKUGQIBELA?

I-Glue ye-Glue ye-Glue ye-Glue ye-Glue ye-Hlues yenziwa phakathi kweziqwenga ezibini zeglasi ezimbini. Kwiglue kakhulu emazantsi, intloko yombhobho ayinakufakwa phakathi kweepleyiti zeglasi ezimbini. Bukela i-glycerin! Beka intloko yombhobho phezu komngxunya wesampulu kwaye isampulu iya kutsiba emngxunyeni.
Isampulu yesampulu ngaphambili, qiniseka ukuba ubeka inqaku elithe nkqo le-Polyproplene Acyl Gel Inzwane inyibilikisiwe icocekile. Hlamba i-acrymide engafakwanga kunye namanzi anokuvela ezantsi komngxunya wesampulu. Amanzi anokwenza isampulu emngxunyeni kakhulu. Sebenzisa i-25ml isirinji okanye i-50ml i-50ml kunye nenaliti ye-18. Thela kwi-elektrophores ye-buffer kwaye uqhume ngononophelo umngxunya wesampulu.
Kunokuba nzima ukubona umngxunya wesampulu, kodwa ngokufanayo, okuseleyo kulula. Ukuba kukho imingxunya engaphezulu, i-buffer yesampulu inokuvavanywa nge-bromophenol iblowu.