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產(chǎn)品詳情
Group:K12
Strain designation:MG1655 SEGA032
Accession number:LMBP 12868
Depositor:A.Ehrmann1 and Prof.Dr M.N?rholm1.It was constructed by C.Bayer1,A.Ehrmann1 and M.Rennig1.
1 The Novo Nordisk Foundation Center for Biosustainability,Technical University of Denmark,Horsholm,Denmark
Containment level:This strain has been assigned the containment level Class 1 following the European Directive 2009/41/EC on the contained use of genetically modified organisms,and its updates(see also the Belgian risk group classification).
Other culture collection numbers:SEGA 032
Medium:LB-Lennox
Selection marker:tetracycline(10μg/ml)
Temperature:37°C
Genotype:F-λ-ilvG-rfb-50 rph-1 glmS-pstS::Ptrc-low-PelB-sfgfp-tetA(Source:http://openwetware.org/wiki/E._coli_genotypes#MG1655+Bayer et al.,2021)
SEGA sequence:p12868.gb
Phenotype:tetR
Properties:The strain was created based on Escherichia coli K12 MG1655,by integrating the SEGA landing pad between the glmS and pstS loci.Counter-selection is posssible through sensitivity to NiCl2(50μM).
Further information:This host strain is intended for simplistic bacterial genome engineering,using the SEGA genome architecture.
Original reference:Bayer et al.,Nat.Commun.12(2021),5876[DOI:10.1038/s41467-021-26155-5]
Restricted use:-BCCM MTA
-The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department.
Refer in your Materials and Methods:
Escherichia coli K12 MG1655 SEGA032(LMBP 12868)is available at BCCM/GeneCorner.It was deposited by A.Ehrmann and Prof.Dr M.N?rholm and was published in Bayer et al.,2021.
常見問題
答:公司提供兩種運輸方式供老師選擇,1、復(fù)蘇的活細胞:采用常溫發(fā)貨的方式,收到即可觀察密度并判斷是否進行傳代操作。優(yōu)勢是省去復(fù)蘇的步驟,細胞成活率較高。2、凍存的細胞:采用干冰運輸,一般情況下發(fā)貨是2支凍存管,收到后放-80過夜,第二天轉(zhuǎn)入液氮長期存儲,擇機復(fù)蘇。優(yōu)勢是發(fā)貨快,一般一兩天即可收到,缺點是需要自己復(fù)蘇。
答:我公司提供的細胞大部分都參考資源庫的培養(yǎng)信息,如ATCC、DSMZ、中科院等等官方平臺。也有少部分細胞為客戶提供了替代培養(yǎng)方案,根據(jù)客戶的意愿進行選擇!
答:不可以重復(fù)使用,一般從我公司發(fā)出的細胞都需要達到一定的密度后發(fā)出,充液的培養(yǎng)基血清比例會比正常培養(yǎng)時所用到的培養(yǎng)液低很多,通常在3-5%,以維持細胞存活,控制生長速度,不可以用來做細胞培養(yǎng)使用。
答:細胞在鏡下發(fā)現(xiàn)圓形的白色的點點,一般情況下是為貼壁的細胞或脫落的細胞死亡后的產(chǎn)物,懸浮細胞也會有這種現(xiàn)象,出現(xiàn)圓形的光圈一樣的圓點。通常,白色的圓點是分散分布的,聚團類的懸浮細胞可能會聚團出現(xiàn)白色的亮斑,技術(shù)老師可以繼續(xù)培養(yǎng)并觀察。
答:一般情況下,我公司建議客戶收到細胞后傳1-2代后即可安排凍存留種,可先凍存1-2支凍存管,凍存的細胞數(shù)量多一些,便于后期復(fù)蘇。購買原代細胞的客戶,要充分考慮該細胞的傳代次數(shù)限制,人源原代細胞大概可以傳7代左右,鼠源的可以傳3代左右,對于一些能傳代次數(shù)很少的原代細胞,不建議凍存,收到后調(diào)整狀態(tài)后即可安排實驗。



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