ʻO ke Kaʻina Exome Ke kanaka holoʻokoʻa
Pono lawelawe
● Māhele Protein Coding Region: ma ka hopu ʻana a me ka hoʻokaʻina ʻana i ka ʻāpana protein coding, hoʻohana ʻia ka hWES e hōʻike i nā ʻano like ʻole e pili ana i ke ʻano protein;
● Pono Kiʻekiʻe: me ka hohonu o ke kaʻina hana kiʻekiʻe, hoʻomaʻamaʻa ka hWES i ka ʻike ʻana i nā ʻano like ʻole a me nā ʻano like ʻole me nā alapine ma lalo o 1%;
● Kūʻai maikaʻi: hWES e loaʻa ma kahi o 85% o ka hoʻololi ʻana i ka maʻi kanaka mai ka 1% o ka genome kanaka;
● ʻElima mau kaʻina hana QC koʻikoʻi e uhi ana i ke kaʻina holoʻokoʻa me Q30>85% hōʻoia.
Hōʻike Laʻana
| Papahana
| Hale Waihona Puke
| Papahana Hopu Exon
| Manaʻo i ka hoʻolālā hoʻonohonoho
|
|
ʻO Illumina NovaSeq Platform
| PE150 | ʻO Agilent SureSelect Human All Exon V6 IDT xGen Exome Hyb Panel V2 | 5 Gb 10 Gb |
Nā Koina Laʻana
| ʻAno Laʻana
| Ka nui(Qubit® )
| Volume
| Hoʻopaʻa ʻana
| Maʻemaʻe (NanoDrop™ ) |
|
DNA Genomic
| ≥ 300 ng | ≥ 15 μL | ≥ 20 ng/μL | OD260/280=1.8-2.0 ʻaʻohe degradation, ʻaʻohe haumia
|
Manaʻo ʻia ʻo Sequencing Depth
No nā maʻi Mendelian/maʻi kakaʻikahi: ʻoi aku ka hohonu o ke kaʻina hana ma luna o 50x
No nā laʻana tumo: ʻoi aku ka hohonu ma luna o 100x
Bioinformatics
Holo Hana Hana
Hāʻawi laʻana
Ka unuhi ʻana o DNA
Kūkulu hale waihona puke
Ke kaʻina ʻana
ʻIkepili ʻikepili
Hāʻawi ʻikepili
Nā lawelawe ma hope o ke kūʻai aku
1. Alignment helu helu
Papa 1 Heluhelu o ka hopena palapala
Papa 2 Heluhelu o ka hopu exome
2. ʻIke Hoʻololi
Helu 1 Heluhelu o SNV a me InDel
3.Kānai Kiʻekiʻe
Kiʻi 2 Circos ʻāpana o Genome-ākea ʻino SNV a me InDel
Papa 3 Ka nānā 'ana i nā genes kumu ma'i
