K-12 MG1655大肠杆菌克隆菌

K-12 MG1655大肠杆菌克隆菌
产品信息：

货号	名称	产品形式	规格	储存
A0042	K-12 MG1655 Strain	甘油菌	300μl	-20℃

 

使用说明：
康朗质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品三个月内通知我司。收到甘油菌后，请及时在固体平板上划线培养，挑取单菌落接种到液体培养基中，震荡培养后使用并保存甘油菌。如需获取其他详细信息请登录我司官网联系QQ客服查询。

基因型：
F- lambda- ilvG- rfb-50 rph-1(NC_000913)

基本信息：
抗性：无
培养基：LB
菌株类别：大肠杆菌
培养条件：37℃，有氧，LB
质粒转化：42℃热激
保存方式：20%甘油，-20℃
基本应用：用于基因克隆
备注：

菌株简介：
K-12 MG1655是一株K系列的大肠杆菌，经检测无Amp和Kan抗性，可用LB培养基，37℃培养，其代谢类型是异养兼性厌氧型，可用于代谢研究、基因编辑等实验。
The original K-12 wild-type strain from the Stanford collection contained both the F plasmid and phage lambda. MG1655 was constructed by curing that strain using acridine orange and UV.This strain was sequenced by the Blattner laboratory because it approximates wild-type E. coli and "has been maintained as a laboratory strain with minimal genetic manipulation, having only been cured of the temperate bacteriophage lambda and F plasmid by means of ultraviolet light and acridine orange, respectively." (Blattner, et al. 1997). The mutations listed in the genotype are present in most K-12 strains and were probably acquired early in the history of the laboratory strain. A frameshift at the end of rph results in decreased pyrE expression and a mild pyrimidine starvation, such that the strain grows 10 to 15% more slowly in pyrimidine-free medium than in medium containing uracil (Jensen 1993). The ilvG- mutation is a frameshift that knocks out acetohydroxy acid synthase II (Lawther, et al. 1982). The rfb-50 mutation is an IS5 insertion that results in the absence of O-antigen synthesis (Liu and Reeves 1994).
MG1655 was derived and named by Mark Guyer from strain W1485, which was derived in Joshua Lederberg's lab from a stab-culture descendant of the original K-12 isolate. This original E. coli strain K-12 was obtained from a stool sample of a diphtheria patient in Palo Alto, CA in 1922 (Bachmann, B., pp. 2460-2488 in Neidhardt et al.1996, Escherichia coli and Salmonella: Cellular and Molecular Biology, ASM Press).MG1655 grows on LB and M9 minimal medium (+ Glucose + 1ug/ml thiamine).

注意事项：
1、为了您的安全和健康，请穿实验服并戴一次性手套操作。本产品仅可用于实验室研究，不能用于动物，人体以及作为食品添加剂等用途。
2、使用甘油菌时可不完全融解，在甘油菌表面蘸取少量涂布固体琼脂平板即可，也可完全融解后使用，但随着冻融次数的增加，菌株的活力会逐渐下降。
 
菌种培养及打管说明：
1、安瓿瓶开封：用浸过75％酒精的脱脂棉擦净安瓿管，用火焰加热其顶端，滴少量无菌水至加热顶端使之破裂，用锉刀或者镊子敲下已破裂的安瓿管顶端。
2、菌株恢复培养：用无菌吸管吸取0.3--0.5ml适宜的液体培养基，滴入安瓿管内，轻轻振荡，使冻干菌体溶解呈悬浮状。吸取全部菌体悬浮液，移植于1-2支建议的培养基试管中，并在建议的条件下培养。
3、注意事项：菌种活化前，请将安瓿管保存在6-10℃的环境下，某些菌种经过冷冻干燥保存后，延迟期较长，需要连续两次继代培养才能正常生长
4、复苏后的菌种在传1-2代后使用。
5、暂不启开的安瓿及复苏后需保藏的斜面应于4℃中保藏。
特别注意事项：
l、微生物菌种应保藏于低温、清洁和干燥的地方,室温放置时问过长会导致菌种衰退;
2、菌种操作应在无菌条件下进行,防止杂菌污染:
3、斜面菌种保藏时间通常为1-2个月,应根据菌种状况及时转接;冻干菌种保藏时间通常为5~10年;
4、菌种使用过程中如出现杂菌污染或菌种生产性能下降，应及时与我司销售联系或更换新的菌种。