2 × Sensi Direct Premix-UNG (Probe qPCR)

Saukewa: HCB5151A

SensiDirect Premix-UNG (Probe qPCR) an tsara shi don yin PCR kai tsaye daga samfurori ba tare da cirewar DNA ba ko shirye-shiryen samfurin.Wannan reagent ya ƙunshi DNA polymerase mai farawa mai zafi, uracil DNA glycosylase (UNG), Mai hana RNase, MgCl.₂, dNTPs (tare da dUTP maimakon dTTP), da stabilizers, don PCR mai ƙididdigewa (qPCR).Wannan reagent preforms high inhibitor-haƙuri, don haka shi za a iya kai tsaye amfani da gano samfurori kamar makogwaro swab, yau, anti-coagulated jini duka, plasma, da kuma serum ba tare da DNA hakar.Reagent yana amfani da buffer na mallakar qPCR tare da gauraye enzymes na anti-inhibitory DNA polymerase da UNG enzyme.Sabili da haka, yana iya samun ingantaccen haɓakar ƙwayoyin da aka yi niyya a cikin samfuran da ke ɗauke da masu hanawa da hana haɓakar haɓakar ƙarya wanda ya haifar da ragowar PCR da gurɓataccen iska.Wannan reagent ya dace da yawancin kayan aikin PCR masu kyalli, kamar Applied Biosystems, Eppendorf, Bio-Rad, Roche da sauransu.

Abubuwan da aka gyara

1. 50× SensiDirect Enzyme/UNG Mix

2. 2× SensiDirect Premix Buffer (dUTP)

Yanayin ajiya

Duk abubuwan da aka gyara yakamata a kiyaye su a -20 ℃ don ajiya na dogon lokaci da 4 ℃ har zuwa watanni 3.Da fatan za a haxa sosai bayan narke da centrifuge kafin amfani.Guji daskarewa akai-akai.

Ka'idar Keke

Umarnin Bututu

1. Ƙarshe na ƙarshe na ƙaddamarwa shine yawanci 0.2μM.Don ingantacciyar sakamako, za'a iya inganta maida hankali a cikin kewayon 0.2-1μM.

2. Gabaɗaya, za'a iya inganta ƙaddamarwar bincike a cikin kewayon 0.1-0.3μM.Mafi kyawun maida hankali na binciken yana da alaƙa da ainihin kayan aikin haɓakawa na PCR, nau'in bincike, da nau'in abun sanya alama mai kyalli.Da fatan za a koma zuwa littafin kayan aiki ko takamaiman buƙatun kowane bincike mai kyalli.

3. Nau'in samfurori daban-daban sun ƙunshi nau'o'in nau'i daban-daban da abun ciki na mai hanawa da kuma kwafin adadin kwayoyin halitta.Ya kamata a yi la'akari da ƙarar samfurin ta ainihin yanayin.Yi dilution na samfurin ta ƙara ruwan da ba shi da ƙwayar cuta ko TE Buffer, idan ya cancanta.

4. Shawarar girma na samfurori daban-daban:

Kula da inganci

1. Ganewar aiki: hankali, ƙayyadaddun ƙayyadaddun da maimaitawa na qPCR.

2. Babu aikin nuclease na waje: babu exogenous endonuclease da exonuclease gurbatawa.

Bayanan kula na Samfur

1. Wannan samfurin yana amfani da sabon nau'in nau'in DNA polymerase mai zafi, wanda za'a iya kunna shi a cikin mintuna 1-5. Tun da an inganta buffer ta amsawa, ya fi dacewa da PCR biyu ko mahara fluorescence ƙididdiga ta amfani da hanyar bincike.

2. Idan darajar Rn na haɓakawa na PCR ya yi ƙasa sosai ko kuma an hana haɓakawa a fili, rage yawan samfurin, ƙara yawan amsawa ko dilution na samfurin na baya zai iya inganta sakamakon.

3. Tarin jini, miya, fitsari, swab, da dai sauransu ya kamata ya bi ka'idodin asibiti, kuma ana iya amfani da sabon samfurin don hana lalatawar acid nucleic.

4. Tun da daban-daban amplicons da daban-daban amfani yadda ya dace zuwa dUTP da hankali ga UNG, da reagents ya kamata a inganta idan ganewa ji na ƙwarai rage lokacin amfani da UNG tsarin.Da fatan za a tuntuɓe mu don tallafin fasaha idan an buƙata.

5. Don guje wa haɓaka samfuran PCR masu ɗaukar nauyi tsakanin halayen mataki ɗaya, ana buƙatar yanki na gwaji da pipette don haɓakawa.Yi aiki da safar hannu kuma canza akai-akai kuma kar a buɗe bututun dauki bayan haɓaka PCR.