PCR Analyzer Troubleshooting: Inowanzo bvunzwa Mibvunzo uye Mhinduro

Polymerase chain reaction (PCR) analyzers maturusi akakosha mumolecular biology, zvichibvumira vaongorori kukudza DNA yezvishandiso kubva pakutsvagisa genetic kusvika kukiriniki yekuongororwa. Nekudaro, senge chero mudziyo wakaoma, PCR analyzer inogona kusangana nematambudziko anokanganisa kuita kwayo. Ichi chinyorwa chinotarisa mimwe mibvunzo inowanzoitika pamusoroPCR analyzerkugadzirisa uye inopa mhinduro dzinoshanda kumatambudziko akajairika.

1. Sei maitiro angu ePCR asiri kuwedzera?

Rimwe rematambudziko anowanzo kutarisana nevashandisi kusakwanisa kwePCR kuita kukwidziridza chinangwa cheDNA. Izvi zvinogona kuverengerwa kune akati wandei zvikonzero:

Zvisizvo primer dhizaini: Ita shuwa kuti maprimers ako akananga kune yakatarwa kutevedzana uye ane yakanyanya kunyungudika tembiricha (Tm). Shandisa maturusi esoftware dhizaini yekutanga kudzivirira kusasungirirana.

Insufficient Template DNA: Ona kuti uri kushandisa yakaringana template DNA. Kunyanya kushoma kunozokonzeresa kusimba kana kusasimba.

Inhibitors mumuenzaniso: Zvinosvibisa mumuenzaniso zvinogona kutadzisa PCR kuita. Funga kuchenesa DNA yako kana kushandisa imwe nzira yekubvisa.

Mhinduro: Tarisa dhizaini yako yekutanga, wedzera template kutarisisa, uye ita shuwa kuti sampuli yako haina inhibitors.

2. Sei PCR yangu chigadzirwa saizi isiriyo?

Kana PCR yako saizi yechigadzirwa isiri sezvaitarisirwa, inogona kuratidza dambudziko nemamiriro ekuita kana zvigadzirwa zvinoshandiswa.

Kwete-chaiyo amplification: Izvi zvinogona kuitika kana primer ikasungirira kune isingatarisirwe saiti. Tarisa uone iwo maprimers uchishandisa chishandiso chakaita seBLAST.

Kusarurama kweAnealing Temperature: Kana tembiricha yekuvharisa yakadzikira zvakanyanya, kusungirirwa kusiri kwakanangana kunogona kuitika. Kukwidziridza tembiricha yekudziya ne gradient PCR.

Solution: Simbisa primer chaiyo uye optimize annealing tembiricha kuti uvandudze huchokwadi hwePCR zvigadzirwa.

3. My PCR analyzer inoratidza meseji yekukanganisa. ndoita sei?

Mharidzo meseji paPCR analyzer inogona kutyisa, asi inogona kupa zviratidzo kune zvinogona kuitika.

Calibration Matambudziko: Ita shuwa kuti PCR analyzer yakarimwa nemazvo. Kugara uchigadzirisa uye cheki cheki kwakakosha kuti uwane mhedzisiro chaiyo.

Software Group: Dzimwe nguva, software bugs inogona kukonzera matambudziko. Tangazve komputa yako uye tarisa kune zvigadziriso zvesoftware.

SOLUTION: Tarisa kune mushandisi bhuku reiyo chaiyo yekukanganisa kodhi uye tevera akakurudzirwa matanho ekugadzirisa matambudziko. Kuchengetedza nguva dzose kunogona kudzivirira matambudziko akawanda.

4. Sei PCR maitiro angu asingaenderane?

Kusawirirana kwePCR mhedzisiro inogona kushungurudza nekuda kwezvikonzero zvakati:

Reagent Hunhu: Ita shuwa kuti ese mareagents, anosanganisira enzymes, buffers, uye dNTPs, matsva uye emhando yepamusoro. Zvinopera kana zvakasvibiswa reagents zvinogona kukonzera kusiyana.

Thermal Cycler Calibration: Isingawirirani tembiricha marongero anogona kukanganisa PCR maitiro. Gara uchitarisa kuenzana kweiyo thermal cycler.

Solution: Shandisa emhando yepamusoro reagents uye gadzirisa yako inopisa cycler nguva nenguva kuti uve nechokwadi chekuenderana.

5. Nzira yekuvandudza sei PCR maitiro ekuita?

Kuvandudza kushanda kwePCR maitiro kunogona kuunza goho repamusoro uye mhedzisiro yakavimbika.

Gadzirisa maitiro ekuita: Edza uchishandisa zvakasiyana-siyana zvekutanga, template DNA uye MgCl2. Yese PCR maitiro angangoda akasiyana mamiriro ekuita kwakaringana.

Shandisa high-fidelity enzymes: Kana kurongeka kwakakosha, funga kushandisa yakakwirira-kutendeseka DNA polymerase kuderedza kukanganisa panguva yekukudza.

Solution: Ita optimization kuyedza kuti uwane akanakisa mamiriro eiyo PCR setup yako.

Muchidimbu

Kugadzirisa matambudziko aPCR analyzerrinogona kunge riri basa rinotyisa, asi kunzwisisa matambudziko akajairika uye mhinduro dzawo kunogona kuwedzera zvakanyanya ruzivo rwako rwePCR. Nekugadzirisa aya matambudziko akajairwa, vaongorori vanogona kuvandudza PCR mhedzisiro uye kuona mhedzisiro yakavimbika mumamorekuru biology application. Kugara uchigadzirisa, kunyatsosarudzwa kwemareagents, uye optimization yemamiriro ekuita ndiwo makiyi ekubudirira kwePCR kuongororwa.