Ulandelelwano lwe-Exome epheleleyo yomntu
Iimpawu zeNkonzo
● Iiphaneli ezimbini ze-exome ezifumanekayo ngokusekelwe ekuphuculeni iithagethi ezineeprobe: i-SureSelect Human All Exon v6 (Agilent) kunye ne-xGen Exome Hybridization Panel v2 (IDT).
- Ngokusekelwe kwiSureSelect Human All Exon V6 Panel, iindawo ze-UTR kunye ne-mtDNA epheleleyo zongezelelwa kuluhlu lokubamba olujoliswe kulo.
- Ukwakhiwa kwelayibrari enikwe amandla yi-UMI kuyafumaneka ngeSureSelect Human All Exon V6 Panel.
● Ukulandelelana kwe-Illumina NovaSeq.
● Umbhobho we-Bioinformatic ojoliswe kuhlalutyo lwesifo okanye kuhlalutyo lwe-tumor.
Iingenelo zeNkonzo
●Iithagethi Ummandla wokubhala iiProteniNgokubamba nokulandelelanisa iindawo zokubhala iiproteni, i-hWES isetyenziselwa ukuveza iintlobo ezahlukeneyo ezinxulumene nolwakhiwo lweproteni.
●Isebenza kakuhle emalini:I-hWES ivelisa malunga ne-85% yotshintsho olunxulumene nesifo somntu oluvela kwi-1% ye-genome yomntu.
●Ukuchaneka Okuphezulu: Ngobunzulu obuphezulu bokulandelelana, i-hWES inceda ekufumaneni zombini iinguqu eziqhelekileyo kunye neenguqu ezingaqhelekanga ezineefrikhwensi ezingaphantsi kwe-1%.
●Ulawulo Oluqinileyo Lomgangatho: Sisebenzisa amanqaku olawulo aphambili amahlanu kuzo zonke izigaba, ukusuka ekulungiseleleni isampuli kunye nethala leencwadi ukuya kulandelelwano kunye ne-bioinformatics. Olu hlolo olucokisekileyo luqinisekisa ukunikezelwa kweziphumo ezisemgangathweni ophezulu rhoqo.
●Uhlalutyo olupheleleyo lweBioinformatics: umbhobho wethu udlula ekuboneni umahluko kwi-genome ebhekisayo, njengoko ubandakanya uhlalutyo oluphambili oluyilelwe ukujongana ngqo nemibuzo yophando enxulumene neempawu zemfuza zezifo okanye uhlalutyo lwethumba.
●Inkxaso emva kokuthengisa:Ukuzibophelela kwethu kudlulela ngaphaya kokugqitywa kweprojekthi kunye nexesha leenkonzo emva kokuthengiswa kweenyanga ezi-3. Ngeli xesha, sinikezela ngokulandela iprojekthi, uncedo lokusombulula iingxaki, kunye neeseshoni ze-Q&A ukuphendula nayiphi na imibuzo enxulumene neziphumo.
Iinkcukacha zeSampuli
| Icebo lokuBamba i-Exon | Icebo Lokulandelelanisa | Iziphumo zeDatha eziCetyisiweyo |
| I-Agilent SureSelectXT Human Zonke i-Exon V6 + UTR + mtDNA I-Agilent SureSelectXT Human All Exon V6 + UTR + mtDNA (ene-UMI) IDT xGen Exome Hyb Panel V2 | I-Illumina NovaSeq PE150 | 5 -10 Gb Kwizifo ze-mendelian/izifo ezingaqhelekanga: > 50x Kwiisampuli zethumba: ≥ 100x |
Iimfuno zeSampuli
| Uhlobo lweSampuli | Isixa(Qubit®) | Ukugxila | Umqulu | Ijeli ye-Agarose |
| I-WES-FF | ≥ 150 ng | ≥ 15 ng/µL |
≥ 15 µL | Akukho okanye ungcoliseko kunye nokuwohloka okulinganiselweyo (Ibhendi ephambili icacile). I-FFPE DNA: Ukuba ibhendi ephambili ayicacanga, kufuneka i-smear eyahlukileyo engaphezulu kwe-2 kb. |
| I-WES-FFPE | ≥ 200 ng | ≥ 15 ng/µL |
I-Bioinformatics
Uhlalutyo lwe-Bioinformatic lweesampuli zesifo se-hWES luquka:
● Ukulandelelanisa idatha QC
● Ulungelelwaniso lweeGenome ezibhekiselele kuzo
● Ukuchongwa kwee-SNP kunye nee-InDels
● Ingcaciso esebenzayo ye-SNPs kunye ne-InDels
Uhlalutyo lwe-Bioinformatic lweesampuli ze-tumor luquka:
● Ukulandelelanisa idatha QC
● Ulungelelwaniso lweeGenome ezibhekiselele kuzo
● Ukuchongwa kwee-SNP, ii-InDels kunye notshintsho lwe-somatic
● Ukuchongwa kweentlobo ezahlukeneyo ze-germline
● Uhlalutyo lweempawu zotshintsho
● Ukuchongwa kwezakhi zofuzo ezibangela uqhushululu ngokusekelwe kwinguquko ye-gain-of-function
● Ingxelo yotshintsho kwinqanaba lokuba umntu usenokuba sengozini yokusebenzisa iziyobisi
● Uhlalutyo lwe-Heterogeneity – ukubalwa kobunyulu kunye ne-ploidy
Ukuhamba komsebenzi wenkonzo
Ukuhanjiswa kwesampulu
Ukukhupha i-DNA
Ukwakhiwa kweThala leeNcwadi
Ukulandelelana
Uhlalutyo lwedatha
Ukuhanjiswa kwedatha
Iinkonzo zasemva kokuthengisa
Idatha QC - Izibalo zokubanjwa kwe-Exome
Ukuchonga umahluko – InDels
Uhlalutyo oluphambili: ukuchongwa nokusasazwa kwee-SNP/InDels eziyingozi – iploti yeCircos
Uhlalutyo lwe-tumor: ukuchongwa kunye nokusasazwa kwe-somatic mutations – Circos plot
Uhlalutyo lwe-tumor: i-clonal lineages
