BMKCloud Log in
条形 banner-03

Lihlahisoa

16S/18S/ITS Amplicon Sequencing-PacBio

Subunit e ho 16S le 18S rRNA e nang le libaka tse bolokiloeng haholo le tse feto-fetohang haholo ke monwana o phethahetseng oa limolek'hule bakeng sa boitsebahatso ba likokoana-hloko tsa prokaryotic le eukaryotic.Ho nka monyetla oa tatellano, li-amplicon tsena li ka shebisisoa ho latela likarolo tse bolokiloeng mme libaka tse feto-fetohang li ka tsebahala ka botlalo bakeng sa boitsebahatso ba likokoana-hloko tse kenyang letsoho lithutong tse akaretsang tlhahlobo ea mefuta-futa ea likokoana-hloko, taxonomy, phylogeny, joalo-joalo. ) tatellano ea sethala sa PacBio se nolofalletsa ho fumana lipalo tse telele tse nepahetseng haholo, tse ka koahelang li-amplicon tsa bolelele bo felletseng (hoo e ka bang 1.5 KB).Pono e pharalletseng ea sebaka sa liphatsa tsa lefutso e matlafalitse haholo tharollo ea tlhaloso ea mefuta ea libaktheria kapa li-fungus.

Sethala:PacBio Sequel II


Lintlha tsa Tšebeletso

Demo Results

Thuto-pale

Melemo ea Tšebeletso

1

● Ho bala nako e telele ho senola tatellano ea bolelele bo felletseng ea 16S/18S/ITS

● Mehala e nepahetseng haholo e nang le tatellano ea mokhoa oa PacBio CCS

● Qeto ea boemo ba mefuta ho tlhaloso ea OTU/ASV

● QIIME2 ea morao-rao e hlahlobang phallo e nang le litlhahlobo tse fapaneng ho latela bobolokelo, litlhaloso, OTU/ASV.

● E sebetsa lithutong tse fapaneng tsa sechaba ka likokoana-hloko

● BMK e na le boiphihlelo bo batsi ka disampole tse fetang 100,000/selemo, e kwahelang mobu, metsi, kgase, seretse, mantle, mala, letlalo, moro o bedisang, dikokonyana, dimela, jj.

● BMKCloud e nolofalitse tlhaloso ea data e nang le lisebelisoa tse 45 tsa ho hlahloba motho ka mong

Litlhaloso tsa Tšebeletso

TatelanoSethala

Laebrari

Lintlha tse khothalelitsoeng

Nako ea ho fetola

PacBio Sequel II

SMRT-tloloko

5K/10K/20K Ditheke

44 Matsatsi a ho sebetsa

Litlhahlobo tsa Bioinformatics

● Taolo ea boleng ba data e tala

● OTU clustering/De-noise(ASV)

● Tlhaloso ea OTU

● Mefuta e sa tšoaneng ea alpha

● Ho fapana ha Beta

● Tlhahlobo ea lihlopha

● Tlhahlobo ea mokhatlo khahlanong le lintlha tsa liteko

● Ho bolela esale pele ka liphatsa tsa lefutso

16sPacbio

Litlhoko tsa Mohlala le Phano

Litlhoko tsa Mohlala:

Bakeng saLintlha tsa DNA:

Mofuta oa Mohlala

Chelete

Ho tsepamisa maikutlo

Bohloeki

Lintlha tsa DNA

> 1 μg

> 20 ng/μl

OD260/280= 1.6-2.5

Bakeng sa mehlala ea tikoloho:

Mofuta oa mohlala

Mokhoa o khothaletsoang oa sampole

Mobu

Chelete ea mohlala: hoo e ka bang.5 g;Lintho tse setseng tse omeletseng li hloka ho tlosoa holimo;Sila likotoana tse kholo 'me u fete ka har'a filthara ea 2 mm;Lisampole tsa Aliquot ka har'a EP-tube kapa cyrotube bakeng sa ho behelloa.

Mantle

Chelete ea mohlala: hoo e ka bang.5 g;Bokella le lisampole tsa aliquot ho EP-tube kapa cryotube e sa sebetseng bakeng sa ho behelloa.

Likahare tsa mala

Mehlala e hloka ho sebetsoa tlas'a boemo ba aseptic.Hlatsoa lithane tse bokeletsoeng ka PBS;Hlakola PBS 'me u bokelle precipitant ka EP-tubes.

Sludge

Chelete ea mohlala: hoo e ka bang.5 g;Bokella le aliquot sludge sampole ho EP-tube e hloekisitsoeng kapa cryotube bakeng sa ho behelloa

Metsi

Bakeng sa sampole e nang le palo e lekanyelitsoeng ea likokoana-hloko, joalo ka metsi a pompo, metsi a seliba, joalo-joalo, Bokella bonyane metsi a 1 L 'me u fete ka 0.22 μm filthara ho ntlafatsa likokoana-hloko tse holim'a lera.Boloka lera ka har'a tube e hloekisitsoeng.

Letlalo

Hlakola bokaholimo ba letlalo ka hloko ka swab e hloekisitsoeng ea k'hothone kapa lehare la ho buoa 'me u le kenye ka har'a tube e sa hloekang.

Tlhahiso ea Mehlala e khothalelitsoeng

Emisa lisampole ka har'a naetrojene e metsi bakeng sa lihora tse 3-4 'me u li boloke ka har'a metsi a naetrojene kapa -80 degree ho boloka nako e telele.Ho hlokahala ho romelloa ka sekepe se nang le leqhoa le omileng.

Phallo ea Mosebetsi oa Ts'ebeletso

tlhahiso ea mohlala

Mohlala oa thomello

Tokiso ea Laebrari

Kaho ea laebrari

Tatelano

Tatelano

Tlhahlobo ea lintlha

Tlhahlobo ea lintlha

Ka mor'a ho rekisa Litšebeletso

Litšebeletso tsa ka mor'a thekiso


  • E fetileng:
  • E 'ngoe:

  • 1.Sekhahla sa tlhaloso ea V3 + V4(Illumina)-based microbial community profiling vs. Full-length (PacBio)-based profiling.
    (Lintlha tsa merero e 30 e khethiloeng ka mokhoa o sa reroang li sebelisitsoe bakeng sa lipalo-palo)

    3

    2. Sekhahla sa tatellano ea amplicon ea bolelele bo felletseng maemong a mefuta ka mefuta e fapaneng ea lisampole.

    4

    3.Kabo ea mefuta

    5
    4.Sefate sa Phylogenetic

    6

    Taba ea BMK

    Ho pepeseha ha arsenic ho baka tšenyo e thibelang mala le ho kenya ts'ebetsong ha sebete sa sebete se lebisang ho ruruha le pyroptosis ea sebete ka matata.

    E hatisitsoe:Saense ea Kakaretso ea Tikoloho,2021

    Leano la tatellano:

    Mehlala: Taolo vs 8 mg / kg sehlopha se pepesitsoeng sa ATO
    Ho lekanya tlhahiso ea data: 102,583 tatelano e tala ea CCS ka kakaretso
    Taolo: 54,518 ± 747 CCS e sebetsang
    ATO-e pepesitsoe : 45,050 ± 1675 CCS e sebetsang

    Liphetho tsa bohlokoa

    Mefuta e fapaneng ea alpha:Ho pepeseha ha ATO ho fetotse haholo bongata ba likokoana-hloko tsa mala le mefuta-futa ea matata.

    Tlhahlobo ea Metastats:
    Boemong ba phylum: 2 bacterial phyla e fumanoa feela ka lihlopha tsa taolo
    Boemong ba leloko: Meloko e 6 e fumanoe e fapane haholo ka bongata
    Boemong ba mefuta: mefuta e 36 e ile ea khetholloa ka kakaretso, 'me e 6 ea eona e fapane haholo ka bongata ba tlhaho.

    Referense

    Thingholm, LB , et al."Batho ba Batenya ba nang le Lefu la tsoekere la Mofuta oa 2 le ba se nang bona ba Bontša Matla le Sebopeho se Fapaneng sa Maleng a Microbial."Cell Host & Microbe26.2(2019).

    fumana qotso

    Ngola molaetsa wa hao mona mme o re romele wona

    Re romelle molaetsa oa hau: