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Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 68717 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 68717-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart |
This material is available to academics and nonprofits only.
Backbone
- Vector backbonepAAV(Search Vector Database)
- Total vector size (bp)7112
- Vector typeAAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)Stbl3
- Growth instructionsDue to ease of recombination, AAV and lentivirus vectors should be amplified in a recombination deficient bacteria strain such as Invitrogen's OneShot Stbl3 cells. Check for integrety of ITR sites with SmaI digest.
- Copy numberLow Copy
Gene/Insert
- Gene/Insert namemRuby2-P2A-GCaMP6s
- SpeciesR. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish)
- Insert Size (bp)2124
- MutationCre-dependent expression from inverted open reading frame (i.e. 'FLEXED')
- PromoterCAG-FLEX
Cloning Information
- Cloning methodUnknown
- 5′ sequencing primeractcagcgctgcctcagtct
- 3′ sequencing primergtttgtacaaatgatgacagcgaag (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- genOway Notice of RIghts
- Industry Terms
- Not Available to Industry
Depositor Comments
mRuby2: Article: Improving FRET dynamic range with bright green and red fluorescent proteins. Lam et al (Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171. PubMed) Addgene Plasmid 40260
GCaMP6s: Article: Ultrasensitive fluorescent proteins for imaging neuronal activity. Chen et al (Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. PubMed) Addgene Plasmid 40753
Information for AAV1 (Catalog # 68717-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6s-WPRE-pA (#68717). In addition to the viral particles, you will also receive purified pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6s-WPRE-pA plasmid DNA.
CAG-driven, Cre dependent, GCaMP6s calcium sensor and bicistronic, physically separate mRuby2 expression. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
- StorageStore at -80℃. Thaw just before use and keep on ice.
- ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
- Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenemRuby2 (Cre-dependent)
Biosafety
Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide
Resource Information
- Terms and Licenses
- Ancillary Agreement for Penn Vectors
- Terms of Use for Viral Vectors
- Industry Terms
- Not Available to Industry
Viral Quality Control
- Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). Thespecific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
- Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.
Visit our viral production page for moreinformation.
Addgene Comments
Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.
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1、有那些品牌的可以选择?我了解的大概有GE、kodak、CRi。还有其他的厂商吗?都有些什么型号。各有什么特点。
2、询价,各个型号的价钱是多少?
3、从敲定到拿到货能用的时间大约是多少?
Luciferase
)
标记细胞或
DNA
,而荧光技术则采用绿色荧光蛋白、红色荧光蛋白等荧光报告基因和
FITC
、
Cy5
、
C
y7
等荧光素及量子点
(quantumdot
,
QD)
进行标记。
小动物活体成像技术是采用高灵敏度制冷
CCD
配合特制的成像暗箱和图像处理软件,使得可以直接监
控活体生物体内的细胞活动和基因行为。实验者借此可以观测活体动物体内肿瘤的生长及转移、
感染性
疾病发展过程、特定基因的表达等生物学过程。
由于具有更高量子效率
CCD
的问世,使活体动物体内光学成像技术具有越来越高的灵敏度,对肿瘤微
小转移灶的检测灵敏度极高;另外,该技术不涉及放射性物质和方法,非常安全。因其操作极其简单、
所得结果直观、
灵敏度高、
实验成本低等特点,
在刚刚发展起来的几年时间内,
已广泛应用于生命科学、
医学研究及药物开发等方面
图2 光声断层成像系统构架图
图3 光声显微镜系统 (a)多轴光纤;(b)聚焦透镜;(c)锥型透镜;(d)光学聚焦镜和超声探测器;(e)水箱;(f)动物固定支架;(g)采样器和参考光纤;(h)温度控制器;(i)电机电源;(j)计算机;(k)数字示波器;(l)控制电机的计算机
目前光声成像的主要研究分支有光声断层成像(Photoacoustic tomography, PAT,见图2)、光声显微成像(Photoacoustic microscopy, PAM,见图3)、光声内窥成像(Intravascular photoacoustic imaging, IVPAI)。光声断层成像清晰地探测到活体小鼠脑血管分布(见图4),根据血容量、血流、血氧等参数反映了脑功能信息。光声成像技术将为脑功能研究提供新的技术手段。基于光声成像反映光吸收的特性,研究者发展了多波长光声成像技术并且应用于肿瘤成像,获得高分辨率的肿瘤新生血管的形态学信息、由血氧饱和度反映的肿瘤代谢信息。光声成像技术为肿瘤的早期诊断与治疗监控提供了强大的技术支持。多波长光声成像在检测活体深层荧光蛋白表达以及基因活性方面取得令人振奋的效果。多波长内窥光声成像针对动脉粥样硬化斑块进行检测,通过光谱解析获得了动脉粥样硬化斑块组份信息(见图5),为光声内窥成像应用于心脑血管疾病检测奠定了实验基础。随着光声显微镜的出现,光声成像发展到了一个新的阶段。光声显微镜将横向分辨率提高了一个数量级达到了45µm。利用光声显微成像技术不仅可以获得高分辨率黑色素瘤的实体和周围的微血管的形态结构图像(见图6),还可以得到活体动物的血氧饱和度信息(见图7)。亚波长光学分辨率光声显微镜的出现将光声成像技术的分辨率提高到前所未有的高度,达到了221 nm。光学分辨率的光声显微镜(OR-PAM)可以轻而易举地对黑色素瘤细胞和血红细胞进行单细胞成像。光声纳米探针的发展为光声成像增添了活力。基于外源光声纳米探针,研究者们发展了光声分子成像和光声治疗。光声分子成像实现了在磁环境中对在血液中循环的肿瘤细胞进行探测以确定肿瘤细胞是否转移,最后发展成了光声流式细胞仪。
图4 光声脑部损伤恢复过程的连续监控成像,(a)~(f)分别为小鼠损伤后第1天、3天、5天、7天、9天和11天的脑部皮层血管光声重建图像;(g)为损伤恢复后小鼠脑部解剖照片
图5 通过光谱解析方法获得的动脉粥样硬化血管内窥光声图像
图6 光声显微镜监测小鼠耳部黑色素瘤生长过程 (a)注射黑色素瘤前小鼠耳部血管的光声显微成像结果;(b)注射部位的在体光声成像,RBC表示红细胞;(c)注射黑色素瘤细胞4天后血管网络光声图像,MT表示黑色素瘤;(d)光学显微成像结果。
图7 使用第二代光学分辨率的显微镜检测老鼠耳朵中血氧含量
作为新一代的无损医学成像技术,光声成像可以无标记地对单个细胞成像、可以对血管形态的高分辨成像、对不同组织的成份进行解析和对血液参数高特异性的功能检测。光声成像实现了从细胞到组织结构的多尺度示踪及功能成像。光声成像可以用于研究动物体脑功能、肿瘤细胞转移和肿瘤形态结构,生理、病理特征,血流异常、药物代谢功能、深层荧光蛋白表达、基因活性等方面的内容,为生物医学应用领域提供了重要研究及监测手段,具有良好的发展前景和广泛的生物医学应用潜力。预期光声成像技术将会引起基础生命科学以及临床医学影像领域的变革。向左转|向右转
不知道哪家单位会有啊?
最近的实验会用到,哪位知道,请不吝赐教,越详细越好,多谢多谢!
如果有,具体在哪个学校/单位/企业?
最近实验需要用到此设备。
如能告诉,不胜感谢!!!谢谢!!!
谢谢!

