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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.

ushicilelo1

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:

ushicilelo1

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

ushicilelo3

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

ushicilelo4

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

ushicilelo5

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

ushicilelo6

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