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Kala-soocidda DNA/RNA - Kala-soocidda Nanopore

Taxanaha DNA/RNA - Taxanaha Nanopore Sawirka Featured

Taxanaha ONT waa hal molecule oo ah tignoolajiyada isku xigxiga calaamadaha korontada ee waqtiga dhabta ah oo ku salaysan nanopores, mabda'a isku xigxiga ee goob kasta waa isku mid. DNA/RNA laba-laaban ayaa ku xidhmi doona borotiinka nanoporos ee ku dhex jira biofilm-ka oo ka nasanaya hogaanka borotiinka matoorka, iyadoo la adeegsanayo farqiga danabka ee labada dhinac ee biofilm-ka, xadhkaha DNA/RNA waxay dhex maraan borotiinka kanaalka nanopores heer go'an. Sababtoo ah kala duwanaanshaha sifooyinka kiimikada ee saldhigyada kala duwan ee xadhigga DNA/RNA, marka hal saldhig ama molecule DNA uu dhex maro kanaalka nanopores, waxay sababi doontaa isbeddelka calaamadaha korantada ee kala duwan. Marka la ogaado oo la waafajiyo calaamadahan, noocyada saldhigga ee u dhigma waa la xisaabin karaa, ogaanshaha waqtiga-dhabta ah ee taxanahana waa la dhammaystiri karaa.

Nala soo xiriir

Faahfaahinta Adeegga

Natiijada Tijaabada

Tilmaamaha Faahfaahinta Adeegga

Madal	Cabbirka Maktabadda	Wax soo saarka Xogta Aragtida (Meelkiiba)	Saxnaanta Hal-saldhig	Codsiyada
Nanopore	8Kb, 10kb, 20kb, Ultralong, cDNA-PCR	70-90Gb/Unug	85-92%	Wicitaanka SV, De novo, Taxanaha dhererka buuxa, Iso-Seq, Faahfaahinta hidda-wadaha, Ogaanshaha methylation-ka DNA-da

Faa'iidooyinka Adeegga

● Khibrad ka badan 5 sano oo ku saabsan madal isku xigxigta PacBio oo leh kumanaan mashruuc oo xiran oo leh noocyo kala duwan.
● BMKGENE waa lammaane rasmi ah oo ka tirsan Oxford Nanopore, oo leh shahaadada laba-geesoodka ah ee RNA/DNA.
● Waxaa jira noocyo caadi ah oo ah qalabka wax lagu sawiro oo leh qalab dhammaystiran iyo wax soo saar ku filan oo isku xigxiga ah.
● Iyada oo lagu saleynayo madal Nanopore, in ka badan 10 cilmi-baaris oo ku saabsan xayawaanka iyo dhirta Denovo ayaa lagu daabacay joornaalada caanka ah ee caalamiga ah.

Shuruudaha Muunadda

Nooca Muunadda

Tirada

Ururinta (Qubit®)

Mugga

Daahirnimo

Kuwa kale

DNA-da hidde-sideyaasha

Ku tiirsanaanta Shuruudaha Xogta

≥20ng/μl

≥15μl

OD260/280=1.7-2.2;

OD260/230≥1.5；

Meel sare oo cad oo ah 260 nm, ma jiraan wax wasakh ah

Xoogga saarista waxaa loo baahan yahay in lagu cabbiro Qubit iyo Qubit/Nanopore ≤ 2

Wadarta RNA-ga

≥1.2μg

≥100μg/μl

≥15μl

OD260/280=1.7-2.5;

OD260/230=0.5-2.5; ma jiraan wax wasakh ah

Qiimaha RIN ≥7.5

Hab-socodka Shaqada Adeegga

Diyaarinta muunadda

Dhismaha maktabadda

Taxanaha

Falanqaynta Xogta

Bixinta mashruuca

Kii hore: Maktabadaha hore loo sameeyay ee Illumina

Xiga: Kala-soocidda Jajabyada La Kordhiyay ee Gaarka ah (SLAF-Seq)

Qiimaynta Tayada Xogta ee Muunadda DNA-da

Shaxda 1. Tirakoobka xogta nadiifka ah.

BMKID	rawSeqNum	rawSumBase	cleanSeqNum	cleanSumBase	nadiifN50Len	nadiifN90Len	cleanMeanLen	cleanMaxLen	cleanMeanQual
DNA_BMK01	1,218,239	26.37	1,121,736	25.90	28,014	15,764	23,090	143,181	9

Qiimaynta Tayada Xogta ee Muunadda RNA

Shaxda 1. Tirakoobka xogta nadiifka ah.

Magaca Faylka	Aqoonsiga Macmiilka	ReadNum	BaseNum	N50	Celceliska Dhererka	Ugu Dheer	Dhibcaha Celceliska ah
RNA_BMK001	C2	8,947,708	4,047,230,083	398	452	129,227	Q12aad