Isesengura rya PCR Gukemura ibibazo: Ibibazo bikunze kubazwa nibisubizo

Isesengura rya polymerase (PCR) ni ibikoresho byingenzi muri biyolojiya ya molekuline, bituma abashakashatsi bongera ADN kubisabwa kuva mubushakashatsi bwerekeranye nubuzima kugeza kwisuzumisha kwa muganga. Ariko, nkibikoresho byose bigoye, isesengura rya PCR rishobora guhura nibibazo bigira ingaruka kumikorere yabyo. Iyi ngingo ikemura ibibazo bimwe bisanzwe bijyanyeIsesengura rya PCRgukemura ibibazo kandi bitanga ibisubizo bifatika kubibazo bisanzwe.

1. Kuki reaction yanjye ya PCR itiyongera?

Kimwe mu bibazo bikunze guhura n’abakoresha ni ukudashobora kwifata kwa PCR kwagura ADN igamije. Ibi birashobora kwitirirwa ibintu byinshi:

Igishushanyo mbonera cya primer kitari cyo: Menya neza ko primers yawe yihariye kurugero rwateganijwe kandi ifite ubushyuhe bwiza bwo gushonga (Tm). Koresha ibikoresho bya software kubishushanyo mbonera kugirango wirinde guhuza bidafite akamaro.

Inyandikorugero idahagije ADN: Menya neza ko ukoresha urugero ruhagije rwa ADN. Guto cyane bizavamo intege nke cyangwa nta amplification.

Inhibitor muri sample: Ibihumanya murugero birashobora kubuza PCR kwitwara. Tekereza kweza ADN cyangwa gukoresha ubundi buryo bwo kuvoma.

Igisubizo: Reba igishushanyo cyawe cya primer, ongera icyitegererezo cyibanze, kandi urebe neza ko icyitegererezo cyawe kitarimo inhibitor.

2. Kuki ibicuruzwa byanjye PCR ari ingano itari yo?

Niba ingano y'ibicuruzwa bya PCR itari nkuko byari byitezwe, irashobora kwerekana ikibazo kijyanye na reaction cyangwa ibikoresho byakoreshejwe.

Kwiyongera kudasanzwe: Ibi birashobora kubaho mugihe primer ihuza urubuga rutateganijwe. Reba umwihariko wa primers ukoresheje igikoresho nka BLAST.

Ubushyuhe bwa Annealing butari bwo: Niba ubushyuhe bwa annealing buri hasi cyane, guhuza bidasanzwe bishobora kuvamo. Gukwirakwiza ubushyuhe bwa annealing na gradient PCR.

Igisubizo: Emeza primer yihariye kandi uhindure ubushyuhe bwa annealing kugirango wongere neza ibicuruzwa bya PCR.

3. Umusesenguzi wanjye PCR yerekana ubutumwa bwamakosa. nkore iki?

Ubutumwa bwibibazo kuri analyseur ya PCR birashobora gutera ubwoba, ariko birashobora gutanga ibimenyetso kubibazo bishobora kuvuka.

Ibibazo bya Calibration: Menya neza ko isesengura rya PCR ryahinduwe neza. Kugenzura buri gihe no kugenzura ni ngombwa kugirango ubone ibisubizo nyabyo.

Itsinda rya software: Rimwe na rimwe, amakosa ya software arashobora gutera ibibazo. Ongera utangire mudasobwa yawe hanyuma urebe niba ivugurura rya software.

SOLUTION: Reba kumfashanyigisho yumukoresha kubijyanye namakosa yihariye hanyuma ukurikize intambwe zisabwa zo gukemura ibibazo. Kubungabunga buri gihe birashobora gukumira ibibazo byinshi.

4. Kuki ibisubizo byanjye bya PCR bidahuye?

Ibisubizo bidahuye bya PCR birashobora gutesha umutwe kubwimpamvu nyinshi:

Ubwiza bwa Reagent: Menya neza ko reagent zose, zirimo enzymes, buffers, na dNTPs, ari shyashya kandi nziza. Reagent yarangiye cyangwa yanduye irashobora gutera guhinduka.

Calibration ya Thermal Cycler: Igenamiterere ridahuye rishobora kugira ingaruka kubikorwa bya PCR. Buri gihe ugenzure kalibrasi ya cycle yumuriro.

Igisubizo: Koresha reagent yo mu rwego rwo hejuru kandi uhindure igare rya buri munsi kugirango umenye ibisubizo bihamye.

5. Nigute dushobora kunoza imikorere ya PCR?

Kunoza imikorere ya PCR irashobora kuganisha kumusaruro mwinshi nibisubizo byizewe.

Hindura uburyo bwo kwitwara: Kugerageza ukoresheje intumbero zitandukanye za primers, icyitegererezo ADN na MgCl2. Buri reaction ya PCR irashobora gusaba ibintu byihariye kugirango bikore neza.

Koresha imisemburo ihanitse cyane: Niba ubunyangamugayo ari ingenzi, tekereza gukoresha polymerase ya ADN yo kwizerwa cyane kugirango ugabanye amakosa mugihe cya amplification.

Igisubizo: Kora igeragezwa ryiza kugirango ubone ibintu byiza bya PCR yihariye.

Muri make

Gukemura ibibazo aIsesengura rya PCRbirashobora kuba umurimo utoroshye, ariko gusobanukirwa ibibazo bisanzwe nibisubizo byabyo birashobora kuzamura cyane uburambe bwa PCR. Mugukemura ibyo bibazo bisanzwe, abashakashatsi barashobora kunoza ibisubizo bya PCR kandi bakemeza ibisubizo byizewe mubikorwa bya biologiya. Kubungabunga buri gihe, guhitamo neza reagent, no gutezimbere imiterere yimikorere ni urufunguzo rwo gusesengura neza PCR.