TOP10感受态细胞
英文名称: TOP10 感受态细胞
型号:null    产品货号: C1200
价格:请致电:010-57128832,18610462672
品牌: 北京

TOP10 感受态细胞说明书
货 号:C1200
规 格:10×100ul / 20×100ul
保 存:-70℃保存,运输为干冰包装。自收货之日起液氮保存至少一年,-70℃保存至少6个月。
产品简介:
本公司生产的TOP10感受态细胞是采用大肠杆菌TOP10菌株经特殊工艺处理得到的感受态细胞,可用于DNA的化学转化。使用pUC19质粒检测,转化效率可达108,-70℃保存六个月转化效率不发生改变。
基因型:F_ mcrAΔ(mrr-hsd RMS-mcrBC) φ80 lacZΔM15△lacⅩ74 recA1 deoR araD139Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG 特 点:一种用于铺制与培养质粒平板和粘粒平板的重级缺陷的抑制型菌株。其中φ80 lacZΔM15基因的产物可与pUC载体编码的β-半乳糖苷酶氨基端实现α-互补,可用于蓝白斑筛选。该菌株适用于高效的DNA克隆和质粒扩增,能保证高拷贝质粒的稳定遗传。 操作方法:(以下各步骤均为无菌操作)
1,将感受态细胞置于冰上融化,以下实验以100ul感受态细胞为例。
2、向感受态细胞悬液中加入需转化的目的DNA,注意目的DNA的体积不要超过感受态细胞悬液体积的十分之一,轻轻旋转离心管以混匀内容物,冰浴放置30分钟。
3、将离心管置于42℃水浴中60-90秒,然后快速转移到冰浴中放置2-3分钟,注意不要摇动离心管。
4、向离心管中加入500ul无菌无抗的SOC或LB培养基,37℃ 180rpm振荡培养1小时。目的是使质粒上相关的抗性标记基因表达,使菌体复苏。
5、取适量已转化的感受态细胞涂布含相应抗生素的SOC或LB平板,37℃倒置培养12-16小时。涂布用量可根据具体实验来调整,如转化的DNA总量较多,可取100ul左右的转化产物涂板;反之,如转化的DNA总量较少,可取200-300ul的转化产物涂板。过多菌液可以抑制细菌生长。如果预计的克隆较少,可通过离心后吸除部分培养液,悬浮菌体后将其涂布于一个平板中。涂布剩余的菌液可4℃保存,如果次日的转化菌落数过少可以将剩下的菌液再涂布新的平板。
注意事项:
1、感受态细胞应保存在-70℃,不可反复冻融,否则其转化效率将会降低。
2、实验过程中应严格无菌操作,防止其它DNA或杂菌的污染,避免为以后的筛选、鉴定带来影响。
3、转化时,转化效率与外源DNA的浓度在一定范围内成正比,但当加入的外源DNA量过多或体积过大反而会降低转化效率。转化时DNA体积要小于感受态细胞体积的十分之一。
4、转化率的计算:转化率=产生菌落的总数/铺板DNA总量。
5、为防止转化实验不成功,可以保留部分连接产物,以重新转化,将损失降到最低。
相关产品:
I1020 IPTG 溶液 (50mg/ml)
A1170 氨苄青霉素储存液(100mg/ml)
K1030 Kanamycin (100mg/ml)卡那霉素
L1015 LB固体培养基(干粉)
L1020 SOC液体培养基(干粉)

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  • 15696-1-AP;HeLa cellHeLa cells were subjected to SDS PAGE followed by western blot with 15696-1-AP(SSRP1 antibody) at dilution of 1:500
  • 15696-1-AP;Ovary cancerImmunohistochemical of paraffin-embedded human ovary tumor using 15696-1-AP(SSRP1 antibody) at dilution of 1:100 (under 40x lens)
  • 15696-1-AP;Ovary cancerImmunohistochemical of paraffin-embedded human ovary tumor using 15696-1-AP(SSRP1 antibody) at dilution of 1:100 (under 10x lens)
  • 15696-1-AP;mouse testis tissueIP Result of anti-SSRP1 (IP:15696-1-AP, 4ug; Detection:15696-1-AP 1:500) with mouse testis tissue lysate 2400ug.
  • 15696-1-AP;HeLa cellsImmunofluorescent analysis of ( -20℃ Ethanol ) fixed HeLa cells using 15696-1-AP( SSRP1 Antibody) at dilution of 1:50 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L)
Positive WB detected in HeLa cells, mouse testis tissue Positive IP detected in Mouse testis tissue Positive IHC detected in Human ovary tumor tissue Positive IF detected in HeLa cells Recommended dilution WB : 1:500-1:5000
IP : 1:200-1:2000
IHC : 1:20-1:200
IF : 1:50-1:500
 SSRP1抗体Application key: WB = Western blotting, IHC = Immunohistochemistry, IF = Immunofluorescence,
IP = Immunoprecipitation
Product Information
Source Rabbit Purification method Antigen affinity purification
Isotype IgG Storage PBS with 0.02% sodium azide and 50% glycerol pH 7.3. Store at -20oC.
Immunogen Information SSRP1抗体
Immunogen SSRP1 fusion protein ag8318 Full name Structure specific recognition protein 1
Calculated molecular
weight
81 kDa Observed molecular 
weight
86 kDa
GenBank accession 
number
BC005116 Gene ID (NCBI) 6749
Gene symbol SSRP1 Synonyms FACT, FACT 80 kDa subunit, FACT complex subunit SSRP1, FACT80, FACTp80, hSSRP1, SSRP1, T160 SSRP1抗体

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