Specific-Locus Amplified Fragment Sequencing (SLAF-Seq)
Kayendedwe kantchito
Technical Scheme
Mawonekedwe a Utumiki
● Kutsata NovaSeq ndi PE150.
● Kukonzekera kwa laibulale yokhala ndi ma barcoding awiri, zomwe zimathandiza kugwirizanitsa zitsanzo zoposa 1000.
● Njira imeneyi ingagwiritsidwe ntchito kapena popanda jini yolozera, ndi mapaipi osiyanasiyana a bioinformatic pa chochitika chilichonse:
Ndi ma genome: Kupezeka kwa SNP ndi InDel
Popanda ma genome: kusanja kwachitsanzo ndi kupezeka kwa SNP
● Mumu silikaMapangidwe a pre-design siteji angapo oletsa ma enzyme ophatikizika amawunikidwa kuti apeze omwe amapanga kugawa kofanana kwa ma tag a SLAF motsatira genome.
● Pakuyesa koyambirira, kuphatikiza kwa ma enzyme atatu kumayesedwa mu zitsanzo za 3 kuti apange malaibulale 9 a SLAF, ndipo chidziwitsochi chimagwiritsidwa ntchito posankha kuphatikiza kwa enzyme yoletsa pulojekitiyi.
Ubwino wa Utumiki
●High Genetic Marker Discovery: Kuphatikiza makina a barcode apamwamba kwambiri amalola kutsatizana kwa anthu ambiri panthawi imodzi, ndipo kukweza kwa malo okhudzana ndi malo kumapangitsa kuti zitheke bwino, kuonetsetsa kuti ma tag akukwaniritsa zofunikira zosiyanasiyana za mafunso osiyanasiyana ofufuza.
● Kudalira Kwambiri pa Genome: Itha kugwiritsidwa ntchito pazamoyo zomwe zili ndi kapena popanda genome.
●Flexible Scheme Design: Single-enzyme, ma enzyme awiri, chimbudzi cha ma enzyme ambiri, ndi mitundu yosiyanasiyana ya michere yonse imatha kusankhidwa kuti ikwaniritse zolinga kapena mitundu yosiyanasiyana ya kafukufuku. Themu silikaKukonzekera koyambirira kumachitika kuti kuwonetsetse kuti ma enzyme apangidwe bwino.
● Kuchita Bwino Kwambiri mu Enzymatic Digestion: Kukonzekera kwa anmu silikakupangiratu komanso kuyesa kutsimikizira kupangidwa koyenera komanso kugawa ma tag a SLAF pa chromosome (1 SLAF tag/4Kb) ndikuchepetsa kubwerezabwereza (<5%).
●Katswiri Wazambiri: Gulu lathu limabweretsa zokumana nazo zambiri pantchito iliyonse, ndi mbiri yotseka ma projekiti opitilira 5000 a SLAF-Seq pa mazana a mitundu, kuphatikiza zomera, zoyamwitsa, mbalame, tizilombo, ndi zamoyo zam'madzi.
● Zodzipangira zokha Bioinformamatic Workflow: BMKGENE idapanga mayendedwe ophatikizika a bioinformatic kwa SLAF-Seq kuti atsimikizire kudalirika komanso kulondola kwa zotulutsa zomaliza.
Mafotokozedwe a Utumiki
| Mtundu wa kusanthula | Chiwerengero cha anthu chovomerezeka | Njira yotsatirira | |
| Kuzama kwa kutsata ma tag | Nambala ya tag | ||
| Genetic Maps | 2 makolo ndi> 150 ana | Makolo: 20x WGS Kuchulukitsa: 10x | Kukula kwa genome: <400 Mb: WGS ndiyofunikira <1Gb: 100K tag 1-2Gb :: 200K ma tag > 2Gb: 300K tag Max 500k ma tag |
| Genome-Wide Association Studies (GWAS) | ≥200 zitsanzo | 10x pa | |
| Kusintha kwa Genetic | ≥30 zitsanzo, ndi> 10 zitsanzo kuchokera gulu lililonse | 10x pa | |
Zofunikira pa Utumiki
Kukhazikika ≥ 5 ng/µL
Chiwerengero chonse ≥ 80 ng
Nanodrop OD260/280=1.6-2.5
Gelisi ya Agarose: palibe kapena kuchepa pang'ono kapena kuipitsidwa
Kutumiza Kwachitsanzo Kovomerezeka
Chidebe: 2 ml centrifuge chubu
(Pazitsanzo zambiri, timalimbikitsa kuti tisasunge ethanol)
Zitsanzo zolembetsera: Zitsanzo ziyenera kulembedwa momveka bwino komanso zofanana ndi zomwe zatumizidwa.
Kutumiza: Owuma-Izi: Zitsanzo ziyenera kulongedzedwa m'matumba kaye ndikukwiriridwa mu ayezi wouma.
Njira ya utumiki
Chitsanzo cha QC
Kuyesera koyendetsa ndege
SLAF-kuyesera
Kukonzekera kwa Library
Kutsata
Kusanthula Zambiri
Pambuyo-kugulitsa Services
Mulinso zowunikira izi:
- Kusintha kwa data QC
- Kukula kwa ma tag a SLAF
Kufotokozera za genome
Popanda genome yofotokozera: kuphatikiza
- Kuwunika kwa ma tag a SLAF.: ziwerengero, kugawa mumtundu wonse
- Kupezeka kwa zilembo: SNP, InDel, SNV, kuyimba kwa CV ndi ndemanga
Kugawidwa kwa ma tag a SLAF pa ma chromosomes:
Kugawa kwa SNPs pa chromosomes:
Chithunzi cha SNP
| Chaka | Journal | IF | Mutu | Mapulogalamu |
| 2022 | Kulankhulana kwachilengedwe | 17.694 | Genomic maziko a giga-chromosomes ndi giga-genome ya mtengo peony Paeonia ostii | SLAF-GWAS |
| 2015 | Phytologist watsopano | 7.433 | Zolemba zapakhomo zimakhazikitsa madera a genomic ofunika kwambiri mu agronomic soya | SLAF-GWAS |
| 2022 | Journal of Advanced Research | 12.822 | Ma genome-wide artificial introgressions of Gossypium barbadense into G. hirsutum wonetsani malo apamwamba kuti muwongolere nthawi imodzi yamtundu wa thonje komanso zokolola makhalidwe | SLAF-Evolutionary genetics |
| 2019 | Chomera cha Molecular | 10.81 | Population Genomic Analysis ndi De Novo Assembly Reveal Origin of Weedy Mpunga ngati Masewera a Chisinthiko | SLAF-Evolutionary genetics |
| 2019 | Nature Genetics | 31.616 | Kutsatizana kwa ma genome ndi mitundu yosiyanasiyana yamtundu wa carp wamba, Cyprinus carpio | Mapu a SLAF-Linkage |
| 2014 | Nature Genetics | 25.455 | Ma genome a mtedza wolimidwa amapereka chidziwitso cha legume karyotypes, polyploid chisinthiko ndi kubzala mbewu. | Mapu a SLAF-Linkage |
| 2022 | Plant Biotechnology Journal | 9.803 | Kuzindikirika kwa ST1 kumawonetsa kusankha komwe kumakhudza kukwera kwa mbeu ndi mafuta ochulukirapo panthawi yoweta soya | Kukula kwa SLAF-Marker |
| 2022 | International Journal of Molecular Sciences | 6.208 | Chizindikiritso ndi DNA Marker Development for Wheat-Leymus mollis 2Ns (2D) Kusintha kwa Disomic Chromosome | Kukula kwa SLAF-Marker |
| Chaka | Journal | IF | Mutu | Mapulogalamu |
| 2023 | Malire mu sayansi ya zomera | 6.735 | Mapu a QTL ndi kusanthula kwapang'onopang'ono kwa shuga panthawi yakucha kwa zipatso za Pyrus pyrifolia | Mapu a Genetic |
| 2022 | Plant Biotechnology Journal | 8.154 | Kuzindikirika kwa ST1 kumawulula kusankha komwe kumakhudza kukwera kwa mbeu ndi kuchuluka kwamafuta pakuweta soya.
| Kuyimba kwa SNP |
| 2022 | Malire mu sayansi ya zomera | 6.623 | Genome-Wide Association Mapu a Hulless Barely Phenotypes mu Chilala Chilengedwe.
| GWAS |
