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Imikhiqizo

Ubude obugcwele

Ngenkathi ukuhlelwa kabusha kwe-MRNA kususelwa kuMnu. Ngakolunye uhlangothi, ukulandelana kwe-pacbio (i-ISO-SEQ) kusebenzisa ubuchwepheshe obufundwa isikhathi eside, okwenza ukulandelwa kokulandelana kwemibhalo emide egcwele. Le ndlela inika amandla okuhlola okuphelele kokunye ukuhlukanisa okuhlukile, ama-gene fusions, kanye ne-poly-adenyylation. Kodwa-ke, kunezinye izinqumo zesichasiso sezichasiso ngenxa yenani eliphakeme ledatha elidingekayo. Ukulungiswa kokulandelana kwePacbio kuncike ku-molecule eyodwa, real-time (SMRT) Ukulandelana, ukuhlinzeka inzuzo ehlukile ekuthumbeni imibhalo emide egcwele. Le ndlela entsha ibandakanya ukusebenzisa ama-zero-mode WaveGuides (ZMWS) kanye nemithombo ye-microfabricated enika amandla ukubonwa kwesikhathi sangempela komsebenzi we-DNA Polymerase ngesikhathi sokulandelana. Ngaphakathi kwalawa ma-ZMW, i-PACBIO's DNA polytherase syntherazes a strand ehambisanayo ye-DNA, edala ukufundwa okude okugcwele okubhaliwe kweMtri. Ukusebenza kwe-PACBIO ekubuyiseleni okulandelanayo kwe-Presensus Sequencing (CCS) kuthuthukisa ukunemba ngokulandelana okulandela i-molecule efanayo. I-HIFI ekhiqizwayo ifundeka inembe okuqhathaniswa nama-NGS, inomthelela phambili ekuhlaziyweni okuphelele nokuthembekile kwezici eziyinkimbinkimbi ze-transcriprymic.

Ipulatifomu: I-Pacbio Sequel II; I-Pacbio Revuo

Xhumana nathi

Imininingwane ye-Service

Bioinformatics

Imiphumela yedemo

Izincwadi Ezifakiwe

Izici

● I-CDNA Synthesis evela ePoly-A Ilandelwa Ukulungiselela Umtapo Welabhulali

● Ukuhlelwa kabusha kumodi ye-CCS, ukukhiqiza ukufundwa kwe-HIFI

● Ukuhlelwa kabusha kwemibhalo emide egcwele

● Ukuhlaziywa akudingi i-genome teferembo; Kodwa-ke, ingaqashwa

● Ukuhlaziywa kwe-bioinformatic kunika amandla ukuhlaziywa kwama-transcript isoform lncrna, gene fusions, poly-adenylation, nesakhiwo sezakhi zofuzo

Izinzuzo zensizakalo

●Ukunemba okuphezulu: I-HIFI ifunda ngokunemba> 99.9% (Q30), iqhathaniswa nama-ngs

● Ukuhlaziywa okuhlukanisayo okuhlukile: Ukulandelana kwayo yonke imibhalo kunika amandla ukukhonjwa kwe-isoform kanye nohluzo

●Ubuchwepheshe obubanzi: Ngerekhodi lokuqedela amaphrojekthi angaphezu kuka-1100 ama-PACBIO agcwele amaphrojekthi futhi acubungule amasampula angaphezu kwama-2300, iqembu lethu liletha umcebo wesipiliyoni kulo lonke iphrojekthi.

●Ukwesekwa ngemuva kokuthengisa: Ukuzibophezela kwethu kufinyelela ngaphezu kokuphothulwa kwephrojekthi ngesikhashana sesevisi esinenyanga esingu-3. Ngalesi sikhathi, sinikezela ngokulandela iphrojekthi, usizo lokuxazulula inkinga, kanye ne-q & amaseshini ukubhekana nanoma yimiphi imibuzo ehlobene nemiphumela.

Izidingo zesampula nokulethwa

Indlu ehlala izincwadi

Ukuhlehlisa Isu

Kunconywe idatha

Ikhwalithi yokulawula

I-Polya icebisa umtapo we-MRNA CCS

I-Pacbio Sequel II

I-Pacbio Revuo

20/40 GB

5/10 m CCS

Q3085%

Izidingo zesampula:

I-nucleotides:

● Izitshalo:

Izimpande, isiqu noma petal: 450 mg

Iqabunga noma Imbewu: 300 mg

Izithelo: 1.2 g

● isilwane:

Inhliziyo noma amathumbu: 300 mg

I-Viscera noma ubuchopho: 240 mg

Imisipha: 450 mg

Amathambo, izinwele noma isikhumba: 1g

● Ama-arthropods:

Izinambuzane: 6G

ICrustacea: 300 mg

● Igazi lonke: 1 ishubhu

● Amaseli: 10⁶amaseli

Conck. (Ng / μL)

Inani (μg)

Ubumsulwa

Ukwethembeka

≥ 100

≥ 1.0

I-OD260 / 280 = 1.7-2.5

I-OD260 / 230 = 0.5-2.5

Kunomkhawulo noma akukho okungcoliswa kweprotheni noma kwe-DNA ekhonjiswe ku-gel.

Wezitshalo: Rin≥7.5;

Okwezilwane: reen8.0;

5.00s 28s / 18S≥1.0;

Kunqunyelwe noma akukho ukuphakama kwesisekelo

Kunconywe ukulethwa kwezinkampu

Isitsha: 2 ml centrifuge tube (i-tin foil ayinconywa)

Ilebula yesampula: Iqembu + liphindaphinda isib. A1, A2, A3; B1, B2, B3.

Ukuthumela:

1. I-Dry-Ice: Amasampula adinga ukupakishwa ezikhwameni futhi angcwatshwa eqhweni eyomile.

2. Amashubhu angenakuzalwa

Ukugeleza komsebenzi wesevisi

Idizayini yokuhlola

Ukulethwa kwesampula

Ukukhishwa kwe-RNA

Ukwakhiwa Kwelabhulali

Ukulandelana

Ukuhlaziywa kwedatha

Services After-Sale

Okwedlule: I-eukaryotic mrna sequenc-ngs

Olandelayo: Ubude obugcwele be-mrna

Kubandakanya ukuhlaziywa okulandelayo:

● Ukulawulwa kwekhwalithi yedatha eluhlaza

● Ukuhlaziywa kwe-polyadenylitation okuhlukile (i-APA)

● Ukuhlaziywa kwe-Fusict Transcript

● Ukuhlaziywa okuhlukanisayo okuhlukile

● Ukulinganisa ama-orthologs we-Universal-Copy Onnerle-Copy (Busco) Ukuhlaziywa

● Ukuhlaziywa kweNoveli Okubhaliwe: Ukubikezela kokulandelanisa kwamakhodi (CD) kanye nesichasiso esisebenzayo

● Ukuhlaziywa kwe-Lncrna: Ukubikezela kweLNCRRA NE-TARGETS

● Ukuhlonza ama-MicroStatelite (SSR)

Ukuhlaziywa kweBusco

Ukuhlaziywa kokuhlukaniswa okuhlukile

Ukuhlaziywa okuhlukile kwe-polyadenylication (APA)

Isichasiselo esisebenzayo semibhalo yenoveli

Hlola intuthuko eyenziwe nge-Bmkgene's Nanopore ebunjiswe ama-nanopore izinsizakalo zokulandelana kwe-Mrna kulolu shicilelo olufakiwe.

Ma, y et al. (2023) 'Ukuhlaziywa kokuqhathanisa kwePacbio kanye ne-Ont RNA Ukuhlehlisa Izindlela ze-Nemopilemabema NoMurai Venem Ukuhlonza', i-genomics, 115 (6), k. 110709. Doi: 10.1016 / J.YGENO.2023.110709.

Chao, Q. et al. . DOI: 10.11111 / PBI.12958.

DEng, H. et al. (2022) Izinguquko ezinamandla kokuqukethwe kwe-ascorbic acid ngesikhathi sokuthuthukiswa kwezithelo nokuvuthwa kwe-Actinidia Latifolia (izitshalo zezithelo ezicebile ezicebile) kanye nezinqubo ze-moleculate ezihlangene ', ama-International Journal of Molecular Science, 23 (10), k. 5808. I-DoI: 10.3390 / IJMS23105808 / S1.

Hua, X. et al. . doi: 10.1038 / s42003-022-03000-Z.

I-LIU, M. et al. (2023) 'Ukuhlaziywa kwe-PACBIO I-ISO-SEQ nokuhlaziywa kwe-tuta absoluta (meyrick) ukuhanjiswa kwe-Tuta Absoluta (meyrick) gescriptipmeme and cytochrome p450 izinhlobo zofuzo, izinambuzane, 14 (4), k. 363. Doi: 10.3390 / Izinambuzane140403633 / S1.

Wang, uLijun et al. . Doi: 10.11186 / S12864-019-5832-9.