Non-Reference Yakavakirwa Mrna Sequencing-NGS
Zvimiro
● Kurwa kwePoly MrN
● Zvakazvimiririra chero renongedzo genome: Kubva kuEf Novo Assangano weManyorero, Kugadzira rondedzero yezvinyorwa izvo zvinonzi madhiri (NR, Swiss-prot, cog, eggnog, pfam, a go, kegg)
● Yakanyatsotemerwa Bioinformatic ongorora yeGene Expression uye transcript chimiro
Zvigadzirwa zvebasa
●Hunyanzvi hwakawanda. Isu takabudirira kuvhara anopfuura zviuru zana emrna-seq mapurojekiti mune akasiyana siyana ekutsvaga nzvimbo.
●Rudzius mhando yekudzoraIzvi Uku kutarisisa kwekutarisa kunovimbisa kuburitsa kwemhedzisiro yemhando yepamusoro yepamusoro.
● Zvakanyatsotaurwa: Isu tinoshandisa madhiri akawanda kuti ashande madhiri anoratidza majini akaratidzwa
●Post-Kutengesa Tsigiro: Kuzvipira kwedu kunowedzera kupfuura projekiti kupedzisa nemwedzi mitatu-mushure-yekutengesa nguva yebasa. Pakati peino nguva, isu tinopa chirongwa chekutevera-up, kugadzirisa dambudziko rekubatsira, uye Q & zvikamu zvekugadzirisa chero mibvunzo zvine chekuita nemhedzisiro.
Mhedzisiro Inodiwa uye Kuendesa
| Raibhurari | Kuongorora zano | Data yakakurudzirwa | Mhando yekudzora |
| Poly yakafumiswa | Illumina Pe150 DNBSEQ-T7 | 6-10 GB | Q3085% |
Zvinodiwa Muenzaniso:
Nucleotide:
| Conc. (Ng / μl) | Mari (μg) | Kuchena | Kuvimbika |
| ≥ 10 | ≥ 0.2 | Od260 / 280 = 1.7-2.5 Od260 / 230 = 0.5-2.5 Limited kana hapana protein kana kusvibiswa kweDNA kunoratidzwa pane gel. | Zvidyarwa: Rin≥4.0; Zvemhuka: Rin≥4.5; 5.0≥28s / 18s≥1.0; Limited kana Hapana Baseline kukwirira |
● Zvirimwa:
Midzi, tsinde kana petal: 450 mg
Shizha kana Mbeu: 300 mg
Michero: 1.2 g
● Mhuka:
Moyo kana mudumbu: 300 mg
Viscera kana uropi: 240 mg
Muswe: 450 mg
Mapfupa, bvudzi kana ganda: 1g
● Arthropood:
Zvipembenene: 6g
Crustacea: 300 mg
● Ropa rese: 1 tube
● Masero: 106 masero
Yakakurudzirwa Samplumple Kutumira
Mudziyo: 2 ml centrifige tube (tin foil haina kukurudzirwa)
Sampula Inonyora: Boka + inodzokorora semuenzaniso A1, A2, A3; B1, B2, B3.
Kutumira:
1. Dry-Ice: samples dzinoda kutariswa mumabhegi uye kuvigwa mune yakaoma-ice.
2..
Basa rebasa rinoyerera
Kuedza Dhizaini
Muenzaniso
Rna kubviswa
Raibhurari Kuvaka
Kutevedzana
Kuongororwa kwedata
Mushure mekutengesa-kutengesa masevhisi
Bioinformatics
TransptemTome Assembly Assembly uye Unigo Selection
UniGene Annotation
Sampula Correlation uye kuongorora kwezvinhu zvakanyorwa zviyedzo
Zvakasiyana-siyana zvinoratidzwa majini (madhiri)
Inoshanda Indiation of DGS
Inoshanda Kufumisa kweDGS
Ongorora kufambira mberi kwakabatana neBMKGECE's Eukaryotic NGRS MrNHER SEQUECE MABHUKU KUNYANYA KUKOSHESA KWEMAHARA eChinyorwa chezvinyorwa.
Shen, F. et Al. (2020) ' Doi: 10.1016 / J.ygeno.2020.01.026.
Zhang, C. et Al. . 212763. DII: 10.3389 / FPLs.016.01425 / bibtex.
Zhu, C. et Al. . Doi: 10.1371 / Shona.pone.0171966.
Zou, l. et al. (2021) ' DOI: 10.1186 / S12870-021-0274-1 / manhamba / 9.
