Highlights
- Simple 20 Second Transformation: No heat shock! Just add DNA and spread on plate.
- High Transformation Efficiencies: Achieve 108 - 109 per µg of plasmid DNA.
- Versatile: Excellent for general cloning and plasmid isolation.
Description
Additional Info | Streptomycin Resistant (StrR) and can not be used for Blue-White Selection. |
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Genotype | F- Δ(gpt-proA)62 leuB6 glnV44 (supE44) ara-14 galK2 lacY1 Δ(mcrC-mrr) xyl-5 mtl-1 recA13 thi-1 rpsL20 (SmR) |
Processing Time | 20 Seconds |
Product Storage | -70°C to -80°C |
Transformation Efficiency | 108 - 109 transformants per µg of plasmid DNA |
Q1: Are the competent cells GMOs?
All our competent cells are classified into Biosafety level 1 and are not genetic modified organisms. Only when transformed with a plasmid they become GMOs.
Q2: Are the Mix & Go! strains dam+ and dcm+?
Most cloning strains will be dam+/dcm+ unless specifically noted in the genotype.
Q3: Do the Mix & Go! strains methylate DNA?
Yes
Q4: Which strains are equivalent to the Zymo strains?
DH5α is equivalent to Zymo 5α. DH10B, Top10, and One Shot Top10 are equivalent to Zymo 10B.For XL-21 Blue, JM109 is the closest match and for Stbl3, HB101 is the closest match.
Q5: How to reduce satellite colonies on agar plates?
– Prepare fresh agar plates– Use more antibiotics in plates– Incubate plates for a shorter time after plating cells
Q6: Is it possible to dilute the competent cells?
We do not recommend diluting the competent cells. We recommend using less DNA to transform cells, or aliquot cells in smaller volumes before transformation. If absolutely necessary, cold 1X Competent Buffer (Mix & Go Transformation Kit, T3001 & T3002) should be used in the dilution.
Q7: Which antibiotics can be used with the Mix & Go! procedure?
No outgrowth is necessary when using Ampicillin or Carbenicillin for selection. However, an outgrowth step is required when using Chloramphenicol, Kanamycin, and Tetracycline because of the mode of action of the antibiotic itself. We recommend the following procedure for the outgrowth step:1. Incubate cells on ice for 5-10 min after addition of plasmid. 2. Add 4 volumes of SOC media.3. Incubate at 37°C for 60 min with gentle shaking at 200-300 rpm.4. Spread on a pre-warmed culture plate containing the appropriate antibiotic.
Q8: Which Plasmid Size can be used for transformation?
For Zymo 5α and Zymo 10B up to 20kb. However, transformation efficiency decreases proportionally from 10-20kb. Above 20kb, cells are difficult to transform. JM109, HB101, XJa, XJa (DE3), XJb, XJb (DE3) and TG1 can handle constructs up to 10kb.
Q9: Which is the recommended DNA concentration and volume for transformation?
There really is no maximum or minimum recommended DNA concentration, but we use 10 pg for quality control. However, the volume of DNA added should not exceed 5% of the cells total volume; the efficiency can decrease several fold as the volume of DNA used increases. If the DNA sample is too diluted, use our DNA Clean & Concentrator.
Q10: What are some tips to improve transformation efficiency?
1. Thaw cells on ice, not room temperature.2. Incubate cells and DNA mixture on ice, not at room temperature. However, do not incubate longer then 1 hour.3. Ensure cells are still frozen when received.4. Pre-warm the culture plates at 37°C for at least 30 minutes.5. Prepare fresh LB agar plates containing the appropriate antibiotic. 6. Prepare a new DNA sample.7. Store the cells at -80°C (not 4°C or -20°C). If the freezer breaks, the cells should be OK as long as the temp does not go higher than -50°C.8. Avoid freeze/thaw cycles.
Q11: How will a heat-shock affect my Transformation Efficiency?
Heat shock is not necessary, however sometimes it can be beneficiary when preparing libraries or transforming XJb Autolysis E. coli strains.We recommend the following protocol for Heat Shock with Outgrowth: 1. Incubate cells on ice for 5-10 min after addition of plasmid. 2. Incubate cells at 42°C for 45 seconds.3. Add 450 ml of SOC to the cells. 4. Incubate at 37°C for 60 min with gentle shaking at 200-300 rpm.5. Spread on a pre-warmed culture plate containing the appropriate antibiotic.
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求助各位大神,点击Downloadfulltable无法下载,总是出现错误,已经排除网络的问题,不知道是否还有其他方法下载,上面有一个Datatable的表格,不知道是不是芯片平台文件?
请问,我通过基因芯片筛选差异基因,有上调的也有下调的,即FC有正值,亦有负值,那么负值怎么取log?谢谢
从GEO和ARRAYEXPRESS下载了miRNA的表达数据矩阵,但是结果不太一样,大致两种:①数值从个位数到几万不等,而且没有小数点;②数值在10上下,有小数点。见图
请问怎么知道数据是否经过了log转换?有注释文件说明吗?还是直接判断①是没转换的,②是转换的?
中唐临界风险,33岁,第一胎,然后羊水穿刺加CMA基因芯片检查,染色体核型正常,4维彩超正常,医生说发育速度偏慢,但还算正常范围,CMA结果异常,如照片
无创dna检测8号染色体缺失32.99mb,羊穿正常,基因芯片检测还是8号染色体杂合性缺失32.2mb,请教专家孩子能要吗?万分感谢
研究蛋白质芯片的意义
1。蛋白质是基因表达的最终产物,接近生命活动的物质层面;
2。探针蛋白特异性高、亲和力强,可简化样品前处理,甚至可直接利用生物材料(血样、尿样、细胞及组织等)进行检测;
3。适合高通量筛选与靶蛋白作用的化合物;
4。有助于了解药物或毒物与其效应相关蛋白质的相互作用。
蛋白质芯片的分类:
1.蛋白质检测芯片
2.蛋白质功能芯片
蛋白质芯片的制备:
1。固相载体及其处理
载体(滴定板、滤膜、凝胶、载玻片)
2。蛋白质的预处理
选择具有较高纯度和完好生物活性的蛋白进行溶解
3。点制微阵列
可使用点制基因微阵列的商品化点样仪或喷墨法等
4。膜为载体:芯片放入湿盒,37°C1h
载玻片为载体:化学修饰产生醛基固定蛋白
5。微阵列的封闭固定微阵列上的蛋白样点
主要封闭试剂:BSA或Gly
相关链接:
全部有关生物芯片的实验方法技术(protocol)
生物芯片相关仪器及芯片・芯片扫描仪・芯片点样仪・生物芯片・生物芯片系统・其它
生物芯片技术服务
核酸分析类试剂
AFLP分析|SNP基因分型|线粒体DNA基因分型|其它基因分型|DNA指纹试剂盒|DNA测序试剂|核酸电泳凝胶|核酸标准品|凝胶纯化试剂盒|核酸染色|转座工具|其它
1、一张芯片上可以同时分析成百上千的探针,这样的情况可以检测多少位点?
2、基因芯片技术通量会有多大?怎么计算?
做蛋白质和microRNA的关系,那么先找出差异的蛋白质还是找出差异的microRNA好呢?有没有做ITRAQ和MICRORNA芯片好的公司推荐,非常感谢!
暂无品牌问答