Babu alamun ƙarawa

1.Taq DNA Polymerase a cikin kit ɗin ya rasa aikinsa saboda rashin ajiya mai kyau ko ƙarewar kit ɗin.
Shawarwari: Tabbatar da yanayin ajiya na kit;sake ƙara adadin da ya dace na Taq DNA Polymerase zuwa tsarin PCR ko siyan sabon Kit ɗin PCR na Real Time don gwaje-gwaje masu alaƙa.

2.Akwai masu hanawa da yawa na Taq DNA Polymerase a cikin samfurin DNA.
Shawara: Gyara samfuri ko rage adadin samfurin da aka yi amfani da shi.

3.The Mg2 + taro bai dace ba.
Shawarwari: Matsakaicin Mg2+ na 2 × Real PCR Mix da muke samarwa shine 3.5mM.Koyaya, ga wasu firamare na musamman da samfura, ƙaddamarwar Mg2+ na iya zama mafi girma.Don haka, zaku iya ƙara MgCl2 kai tsaye don haɓaka taro na Mg2+.Ana ba da shawarar ƙara Mg2+ 0.5mM kowane lokaci don ingantawa.

4.The PCR amplification yanayi ba su dace, da kuma firamare jerin ko maida hankali ne ba daidai ba.
Shawarwari: tabbatar da daidaiton jerin abubuwan farko kuma ba a ƙasƙantar da na'urar ba;idan siginar ƙarawa ba ta da kyau, gwada rage yawan zafin jiki da daidaita ma'auni daidai.

5.Yawan samfuri ya yi kadan ko da yawa.
Shawarwarin: Yi samfuri na daidaitawa gradient dilution, kuma zaɓi ƙaddamarwar samfuri tare da mafi kyawun tasirin PCR don gwajin PCR na Real Time.

NTC yana da ƙimar haske mai girma sosai

1.Reagent gurbatawa lalacewa ta hanyar aiki.
Shawarwari: Sauya tare da sababbin reagents don gwajin PCR na Real Time.

2.Contamination ya faru a lokacin shirye-shiryen tsarin amsawar PCR.
Shawarwari: Ɗauki matakan kariya masu mahimmanci yayin aiki, kamar: sanya safofin hannu na latex, yin amfani da tip ɗin pipette tare da tacewa, da sauransu.

3.The primers suna ƙasƙantar da kai, kuma lalatawar abubuwan da aka yi amfani da su za su haifar da haɓakar da ba ta dace ba.
Shawara: Yi amfani da SDS-PAGE electrophoresis don gano ko abubuwan da aka lalata sun lalace, kuma a maye gurbin su da sabbin maƙallan don gwajin PCR na Real Time.

Dimer na farko ko haɓakawa mara takamaiman

1.The Mg2 + taro bai dace ba.
Shawarwari: Tsarin Mg2+ na 2 × Real PCR EasyTM Mix da muke samarwa shine 3.5 mM.Koyaya, ga wasu firamare na musamman da samfura, ƙaddamarwar Mg2+ na iya zama mafi girma.Don haka, zaku iya ƙara MgCl2 kai tsaye don haɓaka taro na Mg2+.Ana ba da shawarar ƙara Mg2+ 0.5mM kowane lokaci don ingantawa.

2.The PCR annealing zafin jiki ne ma low.
Shawara: Ƙara yawan zafin jiki na PCR da 1℃ ko 2℃ kowane lokaci.

3.A samfurin PCR yayi tsayi da yawa.
Shawarwari: Tsawon samfurin PCR na Real Time yakamata ya kasance tsakanin 100-150bp, bai wuce 500bp ba.

4.The primers suna ƙasƙanci, kuma lalatawar abubuwan da aka tsara za su haifar da bayyanar ƙayyadaddun haɓakawa.
Shawara: Yi amfani da SDS-PAGE electrophoresis don gano ko abubuwan da aka lalata sun lalace, kuma a maye gurbin su da sabbin maƙallan don gwajin PCR na Real Time.

5.Tsarin PCR bai dace ba, ko tsarin ya yi ƙanƙanta.
Shawara: Tsarin amsawar PCR ya yi ƙanƙanta zai sa daidaiton ganowa ya ragu.Zai fi kyau a yi amfani da tsarin amsawa da kayan aikin PCR masu ƙima suka ba da shawarar don sake gudanar da gwajin PCR na Real Time.

Rashin maimaita ƙimar ƙididdiga

1. Kayan aiki yana da matsala.
Shawara: Ana iya samun kurakurai tsakanin kowane rami na PCR na kayan aiki, wanda ke haifar da rashin haɓakawa yayin sarrafa zafin jiki ko ganowa.Da fatan za a bincika bisa ga umarnin kayan aikin da ya dace.

2.Tsarin samfurin ba shi da kyau.
Shawarwari: Samfuran da ba su da tsabta za su haifar da rashin daidaituwa na gwaji, wanda ya haɗa da tsarkin samfuri da masu farawa.Zai fi dacewa don sake tsarkake samfuri, kuma masu farawa sun fi tsarkakewa ta SDS-PAGE.

3.The PCR tsarin shirye-shiryen da lokacin ajiya ya yi tsayi da yawa.
Shawara: Yi amfani da tsarin PCR na ainihi don gwajin PCR nan da nan bayan shiri, kuma kar a bar shi a gefe na dogon lokaci.

4.The PCR amplification yanayi ba su dace, da kuma firamare jerin ko maida hankali ne ba daidai ba.
Shawarwari: tabbatar da daidaiton jerin abubuwan farko kuma ba a ƙasƙantar da na'urar ba;idan siginar ƙarawa ba ta da kyau, gwada rage yawan zafin jiki da daidaita ma'auni daidai.

5.Tsarin PCR bai dace ba, ko tsarin ya yi ƙanƙanta.
Shawara: Tsarin amsawar PCR ya yi ƙanƙanta zai sa daidaiton ganowa ya ragu.Zai fi kyau a yi amfani da tsarin amsawa da kayan aikin PCR masu ƙima suka ba da shawarar don sake gudanar da gwajin PCR na Real Time.