Kodi tiyenera kuganizira chiyani pamene mafayilo a PCR?

Kuti zithandizire bwino, ndikofunikira kuti zigawo zikhalepo zolondola pakukonzekera kulikonse. Kuphatikiza apo, ndikofunikira kuti palibe kuipitsidwa.

Makamaka nthawi zambiri zimapangidwa kuti zikonzedwe kuti zikonzekere kukonzekera zomwe zimasakaniza master m'malo mwapakati pa chotengera chilichonse. Masakani opangidwa kale ndi malonda amapezeka pa malonda, omwe amangotsatira zitsanzo zokhazokha (priger) ndi madzi zimawonjezeredwa. Mwanjira ina, mbuye sakaniza ukhoza kukonzekera nokha. M'matume onse, osakaniza amagawidwa ku chotengera chilichonse cha PCR popanda template ndipo payekha amawonjezeredwa pamapeto pake.

Kugwiritsa ntchito kusakaniza kwa Master kuli ndi zabwino zingapo: Choyamba, kuchuluka kwa masipidwe amodzi kumachepetsedwa. Mwanjira imeneyi, chiopsezo cha zolakwika za ogwiritsa ntchito pakapaimba ndi chiopsezo cha kuipitsidwa chimachepetsedwa ndipo, chabwino, nthawi yasungidwa. Molondola, kulondola kwachiwonetsero kumalinso waukulu, chifukwa mavoliyumu akuluakulu amapangidwa. Izi ndizosavuta kumvetsetsa mukamayang'ana ma piptte a ma pipette: ocheperako osokoneza bongo, zopatuka zimatha. Zowona kuti kukonzekera konse kumachokera ku chombo chomwechi kumathandizanso pa homogeneity (ngati kusakanikirana bwino). Izi zimathandizanso kubereka kwa zoyesazo.

Pokonzekera kusakaniza kwa mbuye, mawu owonjezera 10% ayenera kuwonjezeredwa (mwachitsanzo, kukonzekera 10 kumafunikira, kuwerengera pamaziko a 11), kotero kuti ngakhale chotengera chomaliza chodzazidwa bwino. Mwanjira imeneyi, (pang'ono) pobisalira molakwika, ndipo zotsatira zake zotayirira pamene njira zotchinga zimatha kulipirira. Zotchingira zimapezeka mu enzyme njira monga ma polmerases ndi master sakaniza, ndikupangitsa mafomu a chihemu ndikukhazikika pamtunda wamkatiMalangizo a Pipette.

Kutengera ndi mtundu wamadzi kuti uperekedwe, njira yolondola yaupikisano (1) yosankhidwa ndi zida zoyenera zomwe zasankhidwa. Zothetsera zomwe zili ndi zotchinga, makina oyambitsidwa ndi "chinsinsi chotsika" cha pipette monga njira ina ya mapipu amlengalenga. Zotsatira zaNsonga ya ace pipetteimakhazikitsidwa pazinthu za hydrophobic zapamwamba. Zakumwa zomwe zili ndi zotchinga sizimasiya filimu yotsalira mkati ndi kunja, kotero kuti kutayika kwa yankho kungachepe.

Kuphatikiza pa dosi yokhala ndi zigawo zonse, ndikofunikanso kuti palibe kuipitsidwa kwa kukonzekera kumachitika. Sikokwanira kugwiritsa ntchito zotanuka za chiyero chachikulu, chifukwa njira yopukutira mu pipi la mlengalenga imatha kutulutsa ma aerosols omwe amakhalabe mu pipette. DNA yomwe imapezeka mu aerosol imatha kusamutsidwa kuchokera ku liwiro limodzi kupita ku gawo lotsatirali lotsatira ndikuwongolera kuipitsidwa. Makina osakira mwachindunji omwe atchulidwa pamwambapa atha kuchepetsa chiopsezo ichi. Mapaipi am'mimba amamveka kugwiritsa ntchito malangizo osokoneza bongo kuti ateteze pipette conas, ma aerosols, ndi biosolcul.


Post Nthawi: Desic-06-2022