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產(chǎn)品詳情
Group:K12
Strain designation:MG1655 seqA-mEOS3.2
Accession number:LMBP 9777
Depositor:Prof.Dr J.Michiels1.It was constructed by T.Swings1
1 Centre of Microbial and Plant Genetics,Department of Microbial and Molecular Systems,KU Leuven,Leuven,Belgium
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).
Medium:LB-Miller
Selection marker:kanamycin(50μg/ml)
Temperature:37°C
Genotype:F-λ-ilvG-rfb-50 rph-1 seqA-mEOS3.2:km(R)(Source:http://openwetware.org/wiki/E._coli_genotypes#MG1655+Mika et al.,2015)
Phenotype:KanR
Properties:This strain can be used to study the subcellular localization of the E.coli DNA binding protein SeqA.
Further information:The native seqA gene was fused at its 3'end to the fluorescent protein gene mEOS3.2,separated with a linker(ASPPPGRSR)and followed by a kanamycin resistance marker.The entire cassette was integrated in the chromosome.
Original reference:Mika et al.,Faraday Discuss.184(2015),425-450[PMID:26449690][DOI:10.1039/c5fd00058k]
Restricted use:-BCCM MTA
-The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded,namely UGent-DBMB and VIB-IRC.
Refer in your Materials and Methods:
Escherichia coli K12 MG1655 seqA-mEOS3.2(LMBP 9777)is available at BCCM/GeneCorner.It was deposited by Prof.Dr J.Michiels and was published in Mika et al.,2015.
常見問(wèn)題
答:公司提供兩種運(yùn)輸方式供老師選擇,1、復(fù)蘇的活細(xì)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(jìn)行傳代操作。優(yōu)勢(shì)是省去復(fù)蘇的步驟,細(xì)胞成活率較高。2、凍存的細(xì)胞:采用干冰運(yùn)輸,一般情況下發(fā)貨是2支凍存管,收到后放-80過(guò)夜,第二天轉(zhuǎn)入液氮長(zhǎng)期存儲(chǔ),擇機(jī)復(fù)蘇。優(yōu)勢(shì)是發(fā)貨快,一般一兩天即可收到,缺點(diǎn)是需要自己復(fù)蘇。
答:我公司提供的細(xì)胞大部分都參考資源庫(kù)的培養(yǎng)信息,如ATCC、DSMZ、中科院等等官方平臺(tái)。也有少部分細(xì)胞為客戶提供了替代培養(yǎng)方案,根據(jù)客戶的意愿進(jìn)行選擇!
答:不可以重復(fù)使用,一般從我公司發(fā)出的細(xì)胞都需要達(dá)到一定的密度后發(fā)出,充液的培養(yǎng)基血清比例會(huì)比正常培養(yǎng)時(shí)所用到的培養(yǎng)液低很多,通常在3-5%,以維持細(xì)胞存活,控制生長(zhǎng)速度,不可以用來(lái)做細(xì)胞培養(yǎng)使用。
答:細(xì)胞在鏡下發(fā)現(xiàn)圓形的白色的點(diǎn)點(diǎn),一般情況下是為貼壁的細(xì)胞或脫落的細(xì)胞死亡后的產(chǎn)物,懸浮細(xì)胞也會(huì)有這種現(xiàn)象,出現(xiàn)圓形的光圈一樣的圓點(diǎn)。通常,白色的圓點(diǎn)是分散分布的,聚團(tuán)類的懸浮細(xì)胞可能會(huì)聚團(tuán)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(xì)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(xì)胞數(shù)量多一些,便于后期復(fù)蘇。購(gòu)買原代細(xì)胞的客戶,要充分考慮該細(xì)胞的傳代次數(shù)限制,人源原代細(xì)胞大概可以傳7代左右,鼠源的可以傳3代左右,對(duì)于一些能傳代次數(shù)很少的原代細(xì)胞,不建議凍存,收到后調(diào)整狀態(tài)后即可安排實(shí)驗(yàn)。



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