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Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 62726 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV8 | 62726-AAV8 | Viral service discontinued. | Discontinued |
This material is available to academics and nonprofits only.
Backbone
- Vector backboneAAV-Syn(Search Vector Database)
- Backbone manufactureroriginal from Stratagene
- Backbone sizew/o insert(bp)4368
- Total vector size (bp)6789
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Growth instructionsDH5alpha at 37°C or Stbl3 at 30°C.Carbenicillin is preferred over ampicillin. In DH5alpha this plasmid may act more like a high copy plasmid, although in Stbl3 it may act more like a low copy plasmid.
- Copy numberHigh Copy
Gene/Insert
- Gene/Insert nameChronos-tdTomato
- Alt nameStigeoclonium helveticum channelrhodopsin-tdTomato
- Alt nameChR90-tdTomato
- SpeciesStigeoclonium helveticum
- Insert Size (bp)2421
- GenBank IDKF992040
- Promoterhuman synapsin promoter
- Tag/ Fusion Protein
- tdTomato (C terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBamHI(not destroyed)
- 3′ cloning siteEcoRI(not destroyed)
- 5′ sequencing primergcacgggcgcgaccatctgc
- 3′ sequencing primerTAGCGTAAAAGGAGCAACATAG (Common Sequencing Primers)
Resource Information
- Terms and Licenses
- UBMTA
- Takara Bio Limited Use Label License (formerly Clontech)
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 4 References
Depositor Comments
The plasmid is fully sequenced in the coding sequence regions (opsin-fluorophore and important flanking regions). Multiple digestions were done to verify the vector structure. The construct and the virus were both tested in vitro.
Information for AAV8 (Catalog # 62726-AAV8)(Back to top)
Addgene no longer distributes this item. Contact [email protected] for more information.
Viral service discontinued.
Purpose
Ready-to-use AAV8 particles produced from pAAV-Syn-Chronos-tdTomato (#62726). In addition to the viral particles, you will also receive purified pAAV-Syn-Chronos-tdTomato plasmid DNA.
Syn-driven Chronos-tdTomato expression for optogenetic neural activation.These AAV preparations are suitable purity for injection into animals.Delivery
- Volume.
- Titer≥ 1×10¹³ vg/mL
- Pricing$350 USD for preparation of . 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, AAV8 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV8
- PurificationIodixanol gradient ultracentrifugation
- Reporter GenetdTomato
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.
ebiomall.com






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1903年和丈夫皮埃尔·居里及亨利
直流电流表主要采用磁电系或电动系测量机构(见机械式指示电表测量机构),这些测量机构的测量基本量是电流,可用来直接测小电流。对于大量值的直流电流,磁电系测量机构要使用分流器,也就是并联电阻。它的作用是将大部分被测电流分流。对约10A以下的电流多采用内附分流器;对更大的电流值,则使用专用分流器。它采取四端结构(图a),具有两个电流端,两个电位端。其电阻值的选择条件为:当标称电流通过该分流器时,其电位端间的电压为45mV或75mV;以量程为45mV或75mV的磁电系毫伏表测此电压值,而表盘上则以电流值刻度。对于电
细胞膜离子通道的性质鉴定及其动
力学研究
细胞分泌的研究
信号转导的研究
分子生物学研究向左转|向右转
⒋另外,还应注意仪表的使用环境要符合要求,要远离外磁场。
直流电流表构造
查看图片[电流表]主要包括
三个接线柱[有"+","-"两种接线柱,如(+,-0.6A,-3A)或(-,0.6A,3A)],指针,刻度等(交流电流表无正负接线柱
看你想配什么样的配置了!
我们同事不久前询过一个
好象一套下来是十四万美元左右。
2)内面向外式膜片( inside outpatch)细胞内外和电极内的溶液均可调控,既能较容易地改变细胞内的离子或物质浓度,又能把酶等直接加于膜的内侧面,适宜研究胞内物质对通道活动的影响。但实验中难以改变膜外侧物质,且需浸于低钙液中。常用于研究依赖细胞内钙的离子通道,如钙敏感的钾通道,还可用于细胞内激素和第二信使与通道的调节作用。
3) 外面向外式膜片(outside outpatch)能接触膜的两侧,可以任意改变膜外物质的浓度,有利于研究离子、递质对膜外表面的作用,多用于研究细胞膜外侧受体控制的离子通道。这些受体直接作用于离子通道,而不需经过第二信使系统。因细胞外液容易更换,故加药方便。缺陷是实验中难以改变胞内成分,而且电极管内必须充以低钙液。
4) 全细胞式膜片(whole cell patch)方式使细胞内与浴槽之间的漏流极少。电极本身阻抗(1~10MΩ)与细胞封接后的阻抗相比较低,这种低接触阻抗使单管电压钳容易实现。电极管内与细胞之间弥散交换与平衡快,因而容易控制细胞内液的成分。细胞钳记录的是许多通道的平均电流,有利于综合分析。如果有目的地将膜电位钳制在某一程度,可做到选择性抑制某些通道的活性而只记录某种通道电流的总和,并可在同一细胞上观察几种不同通道的情况。通过改变内部介质,如改变电极液成分,或在电极液中加入所需药物,通过渗透很快改变胞浆成分并达到平衡,该手段在全细胞记录中广泛应用。它适合于小细胞的电压钳位,对于直径大于30μm的细胞很难实现钳位。不足之处是由于电极与细胞间交换快,细胞内环境很容易破坏,因此记录所用的电极液应与胞浆主要成分相同,如高K+,低Na+和Ca2+及一定的缓冲成分和能量代谢所需的物质。
5) 穿孔膜片(perforated patch)是为克服常规全细胞模式的胞质渗漏问题,有学者将与离子亲和的制霉菌素或二性霉素B经微电极灌流到含有类甾醇的细胞膜上,形成只允许一价离子通过的孔,用此法在膜片上做很多导电性孔道,借此对全细胞膜电流进行记录。由于此模式的胞质渗漏极为缓慢,局部串联阻抗较常规全细胞模式高,所以钳制速度很慢,也称为缓慢全细胞模式。
感觉这样的提问没有什么意义
建议,可以自己查阅下资料
用场效应管运算放大器构成的I-V转换器是测量回路的核心部分。在场效应管运算放大器的正负输入端子为等电位,向正输入端子施加指令电位时,由于短路负端子以及膜片都可等电位地达到钳制的目的,当膜片微电极尖端与默片之间形成10GΩ以上封接时,其间的分流电流达到最小,横跨膜片的电流可100%作为来自膜片电极的记录电流(lp)而被测量出来。
这一伟大的贡献,使Neher和Sakmann获得1991年度的诺贝尔生理学与医学奖。
有很多软件可用来处理分析,origin,clampfit,mini analysis·····
电压钳技术是保持细胞膜两侧电压恒定的技术,使细胞膜两次电压稳定在某一个数值,这样就能够测定在这个电位的水平下细胞膜上离子通道的活动情况。
膜片钳技术是固定一片非常小的细胞膜,测定这片膜上的离子通道开放情况,解决了可以单个离子通道活动的测量问题。
现在的膜片钳技术同时包括了以前的电压签技术。

