Iimpawu ze-lncRNA:
1. Ii-lncRNA zidla ngokuba nde, zinentetho eguquguqukayo kunye neendlela ezahlukeneyo zokuhlanganisa ngexesha lokwahlula;
2. Xa kuthelekiswa nohlobo lwekhowudi, i-lncRNA idla ngokuba ngaphantsi;
3. Uninzi lwe-lncRNA luneenkcukacha ezicacileyo zexesha kunye nesithuba kwinkqubo yokwahlulo lwezicubu kunye nophuhliso;
4. Kukho ukubonakaliswa kweempawu kwiithumba kunye nezinye izifo;
5. Iindawo ezisezantsi ze-lncRNA zihlukeneyo;
6. Ulandelelwano luphantsi kulondolozo, kodwa umsebenzi unomlinganiselo othile wolondolozo njalo njalo.
Isisombululo se-LncRNA esibuyisela umva:
Siyazi ukuba i-lncRNA inesakhiwo esikwinqanaba eliphezulu kwiiseli.I-RNA enjalo ihlala inezakhiwo ezinzima ezifana ne-stem loops okanye i-hairpins.Ukuba ulwakhiwo lwesibini lwe-lncRNA alunakusonjululwa ngokufanelekileyo, ukukhutshelwa umva kuthanda ukuba kungaphumeleli okanye kungaphumeleli xa kusenziwa i-RT-qPCR.Ingxaki yokusebenza kakuhle okuphantsi.
Ngokubanzi, kukho amanqaku amathathu aphambili ukuze sifumane itemplates ze-cDNA ezikumgangatho ophezulu:
Umgangatho wethemplethi ye-RNA, iiprimers zokukhuphela umva kunye ne-reverse transcriptase, kunye ne-reverse transcriptase yeyona nto iphambili.I-RNA ehlanjululwe yi-reagents yokutsalwa kwe-RNA esetyenziswa ngokuqhelekileyo ihlala iqulethe iintsalela zotywala kunye neetyuwa ze-guanidine, ezineempembelelo ezinamandla zokuthintela kwizinto ezininzi ze-reverse transcriptases;ukuba elona qondo lobushushu le-reverse transcriptase reaction liphantsi, aliyi kuba luncedo kwi-RNA.Ukuvulwa kolwakhiwo loluhlu.
Okwangoku, iqondo lobushushu elifanelekileyo lereverse transcriptase elisetyenziswa kwiilabhoratri ezininzi lidla ngokuba ngama-42°C, kwaye ubushushu obuphezulu bunokufikelela malunga nama-50°C.Enyanisweni, ukuvulwa kwesakhiwo se-RNA esiphezulu kufuna ubushushu obuphezulu.
Ubushushu bokwenza i-cDNA ye-strand yokuqala ye-Fortune Biotech 2 × RT Easy TM Mix iphezulu njenge-50 ℃-65 ℃, ekulungele ukutyhila ngokupheleleyo isakhiwo esiphezulu se-lncRNA, kwaye ixesha lokusebenza lifutshane.
Ukongeza, imveliso ikwanesakhono esinamandla sokuchasa ukuphazamiseka kwe-RNA kunye nentsalela yotywala kunye netyuwa ye-guanidine, enokuthi ihlangabezane ngokupheleleyo neemfuno zelebhu ukufezekisa impumelelo ephezulu yokukhuphela umva.
Okokuqala Strand cDNA Synthesis Kit
Inombolo yomba: RT-01011 (amaxesha angama-25)
Iimbonakalo:
1. Uzinzo lwe-Thermal: I-65 ℃ isenomsebenzi olungileyo wokukhuphela umva, onceda ukuvula ubume besibini obuntsonkothileyo.
2. Ukunyamezela: i-cDNA synthesis isenokwenziwa ngempumelelo kwiitemplates ze-RNA ngococeko olubi (i-ethanol kunye ne-guanidine iintsalela zetyuwa)
3. Ukukhawuleza: ukusebenza okulula, i-strand yokuqala ye-cDNA synthesis ingagqitywa kwimizuzu engama-25-35.
Usetyenziso olusezantsi: I-PCR, i-qPCR, i-RACE, ukuleyibhelishwa kweetshiphu, ukwakhiwa kwethala leencwadi, njl.
Ukongeza, i-FJ Biotech ibonelela ngeseti epheleleyo yezisombululo ze-RT-PCR ukuhlangabezana neemfuno zeentlobo ezahlukeneyo zabathengi, kubandakanya ezi zixhobo zilandelayo:
IKit yokuQala yeStrand cDNA Synthesis (yeQPCR)
Inombolo yomba: RT-01021 (izihlandlo ezingama-50)
Iimbonakalo:
1. Uncedo: Iiprayimari zokukhuphela umva zongeziwe ukwenza umsebenzi ube lula
2. Uzinzo lwe-Thermal: I-65 ℃ isenomsebenzi olungileyo wokukhuphela umva, onceda ukuvula isakhiwo sesibini esintsonkothileyo.
3. Ukunyamezela: I-cDNA synthesis isenokwenziwa ngempumelelo kwiitemplates ze-RNA ngokungahlambuluki kakuhle
4. Uvakalelo: lufanelekile kwiisampuli ze-RNA eziphantsi
Usetyenziso olusezantsi: Ilungiselelwe ngokukodwa i-qPCR elandelayo
Inyathelo elinye RT-PCR kit
Inombolo yomba: RT-02011 (i-100 amaxesha)
Iimbonakalo:
1. Uzinzo lwe-Thermal: I-65 ℃ isenomsebenzi olungileyo wokukhuphela umva, onceda ukuvula ubume besibini obuntsonkothileyo.
2. Ukunyamezela: I-cDNA synthesis isenokwenziwa ngempumelelo kwiitemplates ze-RNA ngokungahlambuluki kakuhle
3. Ukuchaneka: I-PCR MIX exhotyiswe ngokufanelekileyo iphucula ukusebenza kakuhle kokukhulisa kunye nokuchanekileyo kwendlela yokusabela.
Izicelo ezisezantsi: i-cDNA cloning, uvavanyo lwemfuzo kunye nezinye iimvavanyo ze-molecular
Ikiti ye-RT-PCR enamanyathelo amabini
Inombolo yomba: RT-02021 (amaxesha ali-100)
Iimbonakalo:
1. Ilula kwaye ikhawuleza: ukuphendula ityhubhu enye
2. Uzinzo lwe-Thermal: I-65 ℃ isenomsebenzi olungileyo wokukhuphela umva, onceda ukuvula isakhiwo sesibini esintsonkothileyo.
3. Ukuchaneka: I-Foregene Taq eyongeziweyo iphucula ngempumelelo i-amplification ye-amplification kunye nokucacileyo kwempendulo
Izicelo ezisezantsi: imifuniselo yeemolekyuli ezifana novavanyo lwemfuzo
Inyathelo elinye RT-qPCR kit-SYBR Green I
Inombolo yomba: RT-02111 (amaxesha angama-100)
Iimbonakalo:
1. Elula: impendulo yombhobho omnye
2. Uvakalelo oluphezulu: luyakwazi ukubona iitemplates zekopi ephantsi
3. Ukuhambelana: ilungele uninzi lwezixhobo zobungakanani be-fluorescence
Usetyenziso olusezantsi: ukufunyanwa kweesampulu ze-RNA zokulandelela ezifana neeseli kunye neentsholongwane
Inyathelo elinye RT-qPCR kit-Taqman
Inombolo yomba: RT-02131 (100 amaxesha)
Iimbonakalo:
1. Ngokukhawuleza: fumana i-gene ekujoliswe kuyo
2. Uvakalelo oluphezulu: luyakwazi ukubona iitemplates zekopi ephantsi
3. Ukuhambelana: ilungele uninzi lwezixhobo zobungakanani be-fluorescence
Usetyenziso olusezantsi: ukufunyanwa kweesampulu ze-RNA zokulandelela ezifana neeseli kunye neentsholongwane
I-Forge Bioreverse Transcription Kit inokukunceda ukuba uyisombulule ngcono ingxaki yokukhuphela umva.Kwangaxeshanye, isebenzisana neForge Bio RNA Extraction Kit (enokuthi isuse ngokufanelekileyo iintsalela zeDNA) ukukubonelela ngesisombululo esipheleleyo sovavanyo lweRNA.
Ixesha lokuposa: Aug-27-2021