16S/18S/ITS Amplicon Sequencing-PacBio
Kev Pabcuam Zoo
● Ntev-nyeem nthuav qhia tag nrho-ntev ntu ntawm 16S / 18S / ITS
● Kev hu xov tooj muaj tseeb heev nrog PacBio CCS hom sequencing
● Hom kev daws teeb meem hauv OTU/ASV annotation
● Tseeb QIIME2 tsom xam ntws nrog ntau yam kev soj ntsuam ntawm cov ntaub ntawv, annotation, OTU/ASV.
● Muaj feem xyuam rau ntau hom kev tshawb fawb hauv zej zog microbial
● BMK muaj kev paub dhau los nrog ntau dua 100,000 tus qauv / xyoo, npog av, dej, roj, sludge, quav, hnyuv, tawv nqaij, fermentation broth, kab, nroj tsuag, thiab lwm yam.
● BMKCloud txhawb kev txhais cov ntaub ntawv uas muaj 45 tus kheej cov cuab yeej tshuaj xyuas
Service Specifications
| SequencingPlatform | Lub tsev qiv ntawv | Cov ntaub ntawv pom zoo | Lub sijhawm hloov pauv |
| PacBio Sequel II | SMRT- tswb | 5K/10K/20K Tag | 44 hnub ua haujlwm |
Bioinformatics tsom xam
● Raw cov ntaub ntawv zoo tswj
● OTU pawg/De-noise(ASV)
● OTU cov lus qhia
● Alpha ntau haiv neeg
● Beta ntau haiv neeg
● Kev tsom xam ntawm pab pawg
● Kev txheeb xyuas kev sib koom ua ke tawm tsam qhov kev sim
● Muaj nuj nqi gene kwv yees
Sample Requirements thiab Delivery
Cov Qauv Xav Tau:
RauDNA rho tawm:
| Hom qauv | Tus nqi | Nco ntsoov | Purity |
| DNA rho tawm | ib 1m g | 20 ng / μl | OD 260/280 = 1.6-2.5 |
Rau ib puag ncig cov qauv:
| Hom qauv | Cov txheej txheem kuaj pom pom zoo |
| Av | Sampling tus nqi: approx.5 g yog ';Cov khoom uas tseem tshuav yuav tsum tau muab tshem tawm ntawm qhov chaw;Sib tsoo cov khoom loj thiab hla dhau 2 hli lim;Aliquot kuaj hauv sterile EP-tube lossis cyrotube rau kev tshwj tseg. |
| Cov quav | Sampling tus nqi: approx.5 g yog ';Sau thiab aliquot cov qauv hauv sterile EP-tube lossis cryotube rau kev tshwj tseg. |
| Cov ntsiab lus plab hnyuv | Cov qauv yuav tsum tau ua tiav nyob rau hauv aseptic mob.Ntxuav cov ntaub so ntswg nrog PBS;Centrifuge PBS thiab sau cov dej nag hauv EP-tubes. |
| Sludge | Sampling tus nqi: approx.5 g yog ';Sau thiab aliquot sludge qauv hauv sterile EP-tube lossis cryotube rau kev tshwj tseg |
| Tus dej | Rau cov qauv uas txwv tsis pub muaj microbial, xws li dej kais dej, dej zoo, thiab lwm yam, Sau tsawg kawg yog 1 L dej thiab dhau los ntawm 0.22 μm lim kom txhawb microbial ntawm daim nyias nyias.Khaws cov membrane rau hauv lub raj tsis muaj menyuam. |
| Daim tawv nqaij | Ua tib zoo txhuam cov tawv nqaij nrog cov paj rwb sterile swab los yog phais hniav thiab muab tso rau hauv sterile raj. |
Pom zoo coj mus kuaj
Kho cov qauv hauv cov kua nitrogen rau 3-4 teev thiab khaws cia hauv cov kua nitrogen los yog -80 degree mus rau lub sijhawm ntev.Qauv shipping nrog qhuav-dej khov yog yuav tsum tau.
Kev Pabcuam Ua Haujlwm Flow
Qauv xa tuaj
Kev tsim kho tsev qiv ntawv
Sequencing
Cov ntaub ntawv tsom xam
Kev pab cuam tom qab muag
1.Annotation rate of V3+V4(Illumina)-raws li microbial community profiling vs. Full-length (PacBio)-based profiling.
(Cov ntaub ntawv ntawm 30 randomly xaiv tej yaam num tau thov rau kev txheeb cais)
2.Annotation tus nqi ntawm tag nrho-ntev amplicon sequencing ntawm hom-theem hauv ntau hom qauv
3. Hom kev faib tawm
4.Phylogenetic tsob ntoo
BMK Cas
Kev raug arsenic induces plab hnyuv teeb meem kev puas tsuaj thiab ua rau lub plab-siab axis ua rau o thiab pyroptosis ntawm daim siab nyob rau hauv ducks.
Luam tawm:Kev tshawb fawb ntawm tag nrho ib puag ncig,2021
Sequence Strategy:
Cov Qauv: Tswj vs 8 mg / kg ATO nthuav tawm pawg
Sequencing data yield: 102,583 raw CCS sequences in total
Tswj: 54,518 ± 747 zoo CCS
ATO-exposed: 45,050 ± 1675 zoo CCS
Cov txiaj ntsig tseem ceeb
Alpha ntau haiv neeg:ATO raug cuam tshuam tau hloov pauv txoj hnyuv microbial nplua nuj thiab ntau haiv neeg hauv cov ducks.
Metastats tsom xam:
Hauv qib phylum: 2 kab mob phyla tsuas yog kuaj pom hauv pawg tswj hwm
Hauv qib genus: 6 genera tau pom muaj qhov sib txawv hauv cov txheeb ze ntau
Hauv qib hom: 36 hom tau txheeb xyuas nyob rau hauv tag nrho, nrog rau 6 ntawm lawv sib txawv heev hauv kev nplua nuj
Kev siv
Thingholm, LB, et al."Cov neeg rog rog uas muaj thiab tsis muaj ntshav qab zib hom 2 qhia qhov sib txawv ntawm plab hnyuv microbial muaj peev xwm ua haujlwm thiab muaj peev xwm."Cell Host & Microbe26.2 (2019).
