PacBio 2+3 Yakazara-Kureba mRNA Solution
Features
● Magadzirirwo ezvidzidzo:
Sampuli yakabatanidzwa yakatevedzana nePacBio kuona zvinyorwa zveisoforms
Akapatsanura masampuli (anodzokorora uye mamiriro ekuedzwa) akateedzana nawoNGS kuverengera kutaura kwezvinyorwa
● PacBio sequencing muCCS mode, inogadzira HiFi kuverenga
● Kutevedzana kwezvinyorwa zvehurefu hwakazara
● Kuongorora hakudi referensi genome; zvisinei, inogona kushandiswa
● Ongororo yeBioinformatic haingosanganisire kutaura chete pajene uye isoform-level asiwo kuongororwa kwelncRNA, kusanganiswa kwemajini, poly-adenylation, uye gene chimiro.
Zvakanakira
● Kururama Kwakakwirira: HiFi inoverenga nemazvo> 99.9% (Q30), inofananidzwa neNGS
● Alternative Splicing Analysis: kutevedza zvinyorwa zvese zvinogonesa isoform kuzivikanwa uye hunhu.
● Kusanganiswa kwePacBio uye NGS Strengths: kugonesa kuwanda kwekutaura padanho re isoform, kuvheneka shanduko inogona kuvharika kana uchiongorora iyo yese gene kutaura.
● Unyanzvi Hwakawanda: iine rekodhi yekupedzisa anopfuura zana negumi nemazana ePacBio akazara-kureba transcriptome mapurojekiti uye kugadzirisa anopfuura mazana maviri nemakumi matatu emasampuli, timu yedu inounza hupfumi hweruzivo kupurojekiti yega yega.
● Post-Sales Support: kuzvipira kwedu kunowedzera kupfuura kupera kweprojekiti nemwedzi mitatu mushure mekutengesa sevhisi nguva. Munguva ino, tinopa kutevedza kweprojekiti, rubatsiro rwekugadzirisa matambudziko, uye maQ&A masesheni kugadzirisa chero mibvunzo ine chekuita nemhedzisiro.
Sample Zvinodiwa uye Delivery
| Library | Sequencing strategy | Data inokurudzirwa | Quality Control |
| PolyA yakapfumisa mRNA CCS raibhurari | PacBio Sequel II PacBio Revio | 20/40 Gb 5/10 M CCS | Q30≥85% |
| Poly A yakafumiswa | Illumina PE150 | 6-10 Gb | Q30≥85% |
Nucleotides
|
| Conc.(ng/μl) | Mari (μg) | Kuchena | Kuperera |
| Illumina Library | ≥ 10 | ≥ 0.2 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Yakaganhurirwa kana isina mapuroteni kana kusvibiswa kweDNA kunoratidzwa pagel. | Kumiti: RIN≥4.0; Zvemhuka: RIN≥4.5; 5.0≥28S/18S≥1.0; kukwirira kwakadzika kana kusina kwekutanga |
| PacBio raibhurari | ≥ 100 | ≥ 1.0 | OD260/280=1.7-2.5 OD260/230=0.5-2.5 Yakaganhurirwa kana isina mapuroteni kana kusvibiswa kweDNA kunoratidzwa pagel. | Zvirimwa: RIN≥7.5 Mhuka: RIN≥8.0 5.0≥28S/18S≥1.0; kukwirira kwakadzika kana kusina kwekutanga |
Yakakurudzirwa Sample Delivery
Container: 2 ml centrifuge chubhu (Tin foil haina kukurudzirwa)
Sample labeling: Boka+ replicate eg A1, A2, A3; B1, B2, B3.
Shipment:
1. Dry-aizi:Samples dzinofanirwa kurongedzerwa mumabhegi uye kuvigwa mu dry-ice.
2. RNAstable tubes: RNA samples inogona kuomeswa muRNA stabilization tube (eg RNAstable®) uye inotumirwa mukamuri yekupisa.
Inosanganisira ongororo inotevera:
Raw data quality control
Alternative Polyadenylation Analysis (APA)
Fusion transcript analysis
Alternative Splicing Analysis
Benchmarking Universal Single-Copy Orthologs (BUSCO) ongororo
Novel transcript analysis: kufanotaura kwekutevedzana kwekodhi (CDS) uye chirevo chekushanda
LncRNA kuongororwa: kufanotaura kwelncRNA uye zvinangwa
MicroSatelite Identification (SSR)
Differentially Expressed Transcripts (DETs) kuongorora
Differentially Expressed Genes (DEGs) kuongororwa
Inoshanda annotation yeDEGs uye DETs
BUSCO ongororo
Alternative Splicing Analysis
Alternative Polyadenylation Analysis (APA)
Differentially Akaratidza Genes (DEGs) uye Zvinyorwa (DETs9 anlaysis
Protein-Protein yekudyidzana network yeDETs uye DEGs
Ongorora kufambira mberi kwakafambiswa neBMKGene's PacBio 2+3 yakazara-kureba mRNA inotevedzana kuburikidza neakaunganidzwa muunganidzwa wezvinyorwa.
Chao, Q. et al. (2019) 'Masimba ekusimudzira ePopulus stem transcriptome', Plant Biotechnology Journal, 17 (1), mapeji 206-219. doi: 10.1111/PBI.12958.
Deng, H. et al. (2022) 'Dynamic Changes in Ascorbic Acid Content during Fruit Development and Ripening of Actinidia latifolia (an Ascorbate-Rich Fruit Crop) uye Associated Molecular Mechanisms', International Journal of Molecular Sciences, 23 (10), p. 5808. doi: 10.3390/IJMS23105808/S1.
Hua, X. nevamwe. (2022) 'Kufanotaura kunobudirira kwemajini enzira yebiosynthetic inobatanidzwa mubioactive polyphyllins muParis polyphylla', Communications Biology 2022 5: 1, 5 (1), mapeji 1-10. doi: 10.1038/s42003-022-03000-z.
Liu, M. nevamwe. (2023) 'Yakasanganiswa PacBio Iso-Seq uye Illumina RNA-Seq Kuongorora kweTuta absoluta (Meyrick) Transcriptome uye Cytochrome P450 Genes', Zvipembenene, 14 (4), p. 363. doi: 10.3390/INSECTS14040363/S1.
Wang, Lijun et al. (2019) 'Ongororo ye transcriptome yakaoma uchishandisa PacBio single-molecule chaiyo-nguva yekuongorora yakasanganiswa neIllumina RNA sequencing yekunzwisisa zviri nani ricinoleic acid biosynthesis muRicinus communis', BMC Genomics, 20 (1), mapeji 1-17. doi: 10.1186/S12864-019-5832-9/FIGURES/7.
