-
Kutalika kwathunthu kwa mRNA-Nanopore
Kutsatizana kwa RNA kwakhala chida chamtengo wapatali chowunikira mwatsatanetsatane zolembalemba.Mosakayikira, kutsatizana kwachikale kwa kuwerenga mwachidule kunapindula zambiri zofunika pano.Komabe, nthawi zambiri imakumana ndi zolepheretsa pakuzindikiritsa kwautali wa isoform, kuchuluka, kukondera kwa PCR.
Nanopore sequencing imadzisiyanitsa ndi nsanja zina zotsatizana, chifukwa ma nucleotides amawerengedwa mwachindunji popanda kaphatikizidwe ka DNA ndipo amapanga kuwerengera nthawi yayitali pa makumi a kilobases.Izi zimapereka mphamvu pakuwoloka zolembedwa zazitali zachindunji ndikuthana ndi zovuta mumaphunziro amlingo wa isoform.
nsanja:Nanopore PromethION
Laibulale:cDNA-PCR
-
Kutengera mtundu wautali wa Transcriptome -PacBio
Pa novokutsatizana kwanthawi zonse kwa transcriptome, komwe kumadziwikanso kutiPa novoIso-Seq imatenga ubwino wa PacBio sequencer mu kutalika kwa kuwerenga, zomwe zimathandiza kutsatizana kwa ma molekyulu a cDNA aatali popanda kusweka.Izi zimapewa kwathunthu zolakwika zilizonse zomwe zimapangidwa pamasitepe amisonkhano yolembera ndikupanga ma seti a unigene okhala ndi isoform-level resolution.Ma seti a unigenewa amapereka chidziwitso champhamvu cha majini monga "reference genome" pa transcriptome-level.Kuphatikiza apo, kuphatikiza ndi deta yotsatizana ya m'badwo wotsatira, ntchitoyi imapatsa mphamvu kuwerengera kolondola kwa mawu amtundu wa isoform.
Pulatifomu: PacBio Sequel IILibrary: SMRT belu laibulale -
Eukaryotic mRNA sequencing-Illumina
Kutsatizana kwa mRNA kumathandizira kuti mbiri ya mRNA yonse yolembedwa kuchokera ku cell ikhale yodziwika.Ndi ukadaulo wamphamvu wowulula mbiri ya jini, kapangidwe ka majini ndi mamolekyu azinthu zina zachilengedwe.Mpaka pano, kutsatizana kwa mRNA kwagwiritsidwa ntchito kwambiri pakufufuza kofunikira, zowunikira zamankhwala, chitukuko chamankhwala, ndi zina zambiri.
Platform: Illumina NovaSeq 6000
-
Non-Reference based mRNA sequencing-Illumina
Kutsatizana kwa mRNA kumatengera njira yotsatirira ya m'badwo wotsatira (NGS) kuti ijambule messenger RNA(mRNA) mawonekedwe a Eukaryote panthawi inayake yomwe ntchito zina zapadera zikuyatsidwa.Zolemba zazitali kwambiri zomwe zidapangidwa zimatchedwa 'Unigene' ndipo zimagwiritsidwa ntchito ngati njira yotsatirira pakuwunikiridwa kotsatira, yomwe ndi njira yabwino yophunzirira momwe maselo amagwirira ntchito komanso maulamuliro amtundu wamtunduwu popanda kutchulidwa.
Pambuyo pa kusonkhanitsa deta ya transcriptome ndi kumasulira kwa unigene
(1) Kusanthula kwa SNP, kusanthula kwa SSR, kuneneratu kwa CDS ndi kapangidwe ka jini zidzakonzedweratu.
(2) Kuchulukitsa kwa mawu a unigene mu chitsanzo chilichonse kudzachitika.
(3) Ma unigene omwe amawonetsedwa mosiyanasiyana pakati pa zitsanzo (kapena magulu) apezeka potengera mawonekedwe
(4) Kuphatikizika, kutanthauzira kogwira ntchito ndi kusanthula kopindulitsa kwa ma unigene ofotokozedwa mosiyanasiyana kudzachitika
-
Kutsata kwanthawi yayitali kosalemba-Illumina
Ma RNA autali osalemba zilembo (lncRNAs) ndi mtundu wa mamolekyu a RNA okhala ndi kutalika kopitilira 200 nt, omwe amadziwika ndi kuthekera kotsika kwambiri.LncRNA, monga membala wofunikira mu ma RNA osalemba ma code, amapezeka makamaka mu nucleus ndi plasma.Kukula kwa ukadaulo wotsatizana ndi bioinformatics kumathandizira kuzindikira ma LncRNA angapo ndikuphatikiza omwe ali ndi ntchito zachilengedwe.Umboni wochuluka umasonyeza kuti lncRNA imakhudzidwa kwambiri ndi epigenetic regulation, transcript regulation ndi post-transcript regulation.
-
Small RNA sequencing-Illumina
RNA yaying'ono imatanthawuza gulu la ma molekyulu a RNA osalemba zilembo omwe nthawi zambiri amakhala osakwana 200nt m'litali, kuphatikiza yaying'ono RNA (miRNA), kusokoneza pang'ono RNA (siRNA), ndi piwi-interacting RNA (piRNA).
MicroRNA (miRNA) ndi gulu la endogenous RNA yaying'ono yokhala ndi kutalika kwa pafupifupi 20-24nt, yomwe imagwira ntchito zingapo zofunika zowongolera m'maselo.miRNA imakhudzidwa ndi zochitika zambiri zamoyo zomwe zimawulula minofu - yeniyeni ndi siteji - kufotokozera komanso kusungidwa kwambiri mumitundu yosiyanasiyana.
-
circRNA sequencing-Illumina
Masewero amtundu wonse wa transcriptome adapangidwa kuti aziwonetsa mitundu yonse ya mamolekyu a RNA, kuphatikiza ma coding (mRNA) ndi ma RNA osalemba (kuphatikiza lncRNA, circRNA ndi miRNA) omwe amalembedwa ndi ma cell enieni panthawi inayake.Kutsata kwamtundu wonse, komwe kumadziwikanso kuti "total RNA sequencing" ikufuna kuwulula maulamuliro athunthu pamlingo wa transcriptome.Kutengera mwayi paukadaulo wa NGS, kutsatizana kwazinthu zonse za transcriptome kulipo pakuwunika kwa ceRNA ndikuwunika kwa RNA, komwe kumapereka gawo loyamba lodziwika bwino.Kuwulula maukonde owongolera a circRNA-miRNA-mRNA based ceRNA.
-
Kutsatizana konse kwa transcriptome - Illumina
Masewero amtundu wonse wa transcriptome adapangidwa kuti aziwonetsa mitundu yonse ya mamolekyu a RNA, kuphatikiza ma coding (mRNA) ndi ma RNA osalemba (kuphatikiza lncRNA, circRNA ndi miRNA) omwe amalembedwa ndi ma cell enieni panthawi inayake.Kutsata kwamtundu wonse, komwe kumadziwikanso kuti "total RNA sequencing" ikufuna kuwulula maulamuliro athunthu pamlingo wa transcriptome.Kutengera mwayi paukadaulo wa NGS, kutsatizana kwazinthu zonse za transcriptome kulipo pakuwunika kwa ceRNA ndikuwunika kwa RNA, komwe kumapereka gawo loyamba lodziwika bwino.Kuwulula maukonde owongolera a circRNA-miRNA-mRNA based ceRNA.
-
Kutsata kwa Prokaryotic RNA
Kutsatizana kwa Prokaryotic RNA kumagwiritsa ntchito kutsata kwa m'badwo wotsatira (NGS) kuwulula kupezeka ndi kuchuluka kwa RNA panthawi inayake, posanthula kusintha kwa ma transcriptome a cell.Tsatanetsatane wa kampani yathu ya prokaryotic RNA, makamaka imayang'ana ma prokaryotes okhala ndi ma genome, kukupatsirani mbiri ya transcriptome, kusanthula kapangidwe ka jini, ndi zina zambiri. Zagwiritsidwa ntchito kwambiri pakufufuza koyambira kwa sayansi, kafukufuku wamankhwala ndi chitukuko, ndi zina zambiri.
Platform: Illumina NovaSeq 6000
-
Kutsata kwa Metatranscriptome
Kutsatizana kwa metatranscriptome kumazindikiritsa ma jini a ma microbes (onse a eukaryotes ndi ma prokaryotes) mkati mwa chilengedwe (ie nthaka, madzi, nyanja, ndowe, ndi m'matumbo). za mitundu, kusanthula magwiridwe antchito amitundu yosiyanasiyana, ndi zina zambiri.
Platform: Illumina NovaSeq 6000
