
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 100841 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 100841-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV5 | 100841-AAV5 | Virus (100 µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV9 | 100841-AAV9 | 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)
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Copy numberUnknown
Gene/Insert
- Gene/Insert nameGCaMP6m
- Alt nameGCaMP3 variant 632
- SpeciesSynthetic
- Insert Size (bp)1353
- MutationGCaMP3-T302L R303P M378G K379S D380Y T381R S383T R392G
- PromoterSynapsin
Cloning Information
- Cloning methodUnknown
- 5′ sequencing primerunknown (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- pAAV.Syn.GCaMP6m.WPRE.SV40 (p2823)_Q.gb
- Terms and Licenses
- UBMTA
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 2 References
Depositor Comments
This plasmid was previously available as pAAV.Syn.GCaMP6m.WPRE.SV40 (p2823) from the Penn Vector Core. This plasmid was created as part of the GENIE project at Janelia Research Campus.
Information for AAV1 (Catalog # 100841-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV.Syn.GCaMP6m.WPRE.SV40 (#100841). In addition to the viral particles, you will also receive purified pAAV.Syn.GCaMP6m.WPRE.SV40 plasmid DNA.
Syn-driven GCaMP6m calcium sensor. 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
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.
Information for AAV5 (Catalog # 100841-AAV5)(Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV.Syn.GCaMP6m.WPRE.SV40 (#100841). In addition to the viral particles, you will also receive purified pAAV.Syn.GCaMP6m.WPRE.SV40 plasmid DNA.
Syn-driven GCaMP6m calcium sensor. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 7×10¹² 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, AAV5 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV5
- PurificationIodixanol gradient ultracentrifugation
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
- 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.
Information for AAV9 (Catalog # 100841-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from pAAV.Syn.GCaMP6m.WPRE.SV40 (#100841). In addition to the viral particles, you will also receive purified pAAV.Syn.GCaMP6m.WPRE.SV40 plasmid DNA.
Syn-driven GCaMP6m calcium sensor. 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, AAV9 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV9
- PurificationIodixanol gradient ultracentrifugation
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.
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本人新入植物化学,实验上遇到了一些问题,还望各位大神帮帮忙。
之前通过薄层色谱确定了最佳的展开剂以及展开剂的最佳比例,下一步将过硅胶柱。但是想通过将分光光度计与柱层析联用,将接收液直接进行紫外吸收,以此确定接收的液体为同一物质。但是现在遇到的问题是,在实验过程中是只需要用一种波长去测定还是要不停地调整?(现在只知道物质的极性,不知道组分中有哪些物质)
另外,是通过这种方法分离好一些还是过了柱子后再进行一次薄层色谱好一些呢?
各位大神帮帮忙,感激不尽!!!
分光光度计已经成为现代分子生物实验室常规仪器。常用于核酸,蛋白定量以及细菌生长浓度的定量。
仪器主要由光源、单色器、样品室、检测器、信号处理器和显示与存储系统组成。
2、以空白为参比(或水)测得系列吸光度;
3、以标准溶液的吸光度为纵坐标,对应的标准溶液的含量为横坐标,找出相对的点;
4、将点用一条直线连接起来;尽量将点都落在线上;(绘制标准曲线;)
5、由此根据测得的样品的吸光度查找相对的含量;
1,被测液浓度过高造成透过样品的光信号检测不到,这种情况下可以通过稀释样品来完成;
2,测定时,样品室内的样品架在移动时没移动到位造成挡光;
3,仪器内部的光源不亮或光源没有进入单色器中。可以对照以上情况检查分析!
①辐射源。必须具有稳定的、有足够输出功率的、能提供仪器使用波段的连续光谱,如钨灯、卤钨灯(波长范围350~2500纳米),氘灯或氢灯(180~460纳米),或可调谐染料激光光源等。
②单色器 。它由入射、出射狭缝、透镜系统和色散元件(棱镜或光栅)组成,是用以产生高纯度单色光束的装置,其功能包括将光源产生的复合光分解为单色光和分出所需的单色光束。
③试样容器,又称吸收池。供盛放试液进行吸光度测量之用,分为石英池和玻璃池两种,前者适用于紫外到可见区,后者只适用于可见区。容器的光程一般为0.5~10厘米。
④检测器,又称光电转换器。常用的有光电管或光电倍增管,后者较前者更灵敏,特别适用于检测较弱的辐射。近年来还使用光导摄像管或光电二极管矩阵作检测器,具有快速扫描的特点。
⑤显示装置。这部分装置发展较快。较高级的光度计,常备有微处理机、荧光屏显示和记录仪等,可将图谱、数据和操作条件都显示出来。
①辐射源。必须具有稳定的、有足够输出功率的、能提供仪器使用波段的连续光谱,如钨灯、卤钨灯(波长范围350~2500纳米),氘灯或氢灯(180~460纳米),或可调谐染料激光光源等。
②单色器 。它由入射、出射狭缝、透镜系统和色散元件(棱镜或光栅)组成,是用以产生高纯度单色光束的装置,其功能包括将光源产生的复合光分解为单色光和分出所需的单色光束。
③试样容器,又称吸收池。供盛放试液进行吸光度测量之用,分为石英池和玻璃池两种,前者适用于紫外到可见区,后者只适用于可见区。容器的光程一般为0.5~10厘米。
④检测器,又称光电转换器。常用的有光电管或光电倍增管,后者较前者更灵敏,特别适用于检测较弱的辐射。近年来还使用光导摄像管或光电二极管矩阵作检测器,具有快速扫描的特点。
⑤显示装置。这部分装置发展较快。较高级的光度计,常备有微处理机、荧光屏显示和记录仪等,可将图谱、数据和操作条件都显示出来。
本人实验新人一枚,现在做的是PCR后内切来验证基因多态性,最近遇到了一个难以解决的问题。
本人现在现在用的是天跟的PCRMIX,然后用的是omega的纯化试剂,现在遇到的问题是:我纯化后的PCR产物(只有244bp),在酶标仪、分光光度计检测下浓度相差不大,都是50ul的PCR体系中可以提取出体积35ul,浓度约50-60ng/UL的,纯度1.8-2.0之间纯化液,但是电泳看不到除了Marker之外的条带(图一),即使我加DNA总量达到了几百ng,仍然是什么都看不见,但是似乎隐约有点条带,联系OMEGA的技术后考虑其实并没有回收那么多的DNA,建议我加用异丙醇,但是我加了几百ng都看不见,我觉得加用异丙醇意义不是特别大,而且我是要做内切酶的,对DNA是有要求的(1ug),如果他们的试剂真的存在干扰浓度测定的话,接下来内切我也不好做,所以来求助一下各位大神,是否实验还有所改进,还是更换一些能更好提取小分子DNA的试剂。
PS:我做了引物浓度(华大基因)、模板DNA浓度、Tm的梯度,发现我的模板DNA需要加到几百ng,引物浓度需要加到1.6uM,才能得到比较清晰的条带,但是引物二聚体(40bp)在琼脂糖上一直很明显(图二,目的条带看起来不明显的原因是手机没拍好),PCR的纯化产物也应该是目的条带,因为我在不同引物浓度下的PCR,引物二聚体的条带亮度差别较大,但是纯化产物在酶标仪、分光光度计检测下浓度相差不大,所以应该是产物浓度。现在也是一筹莫展,**各位大神指点。
我之前好像也在丁香园看到过说是部分试剂存在这种情况,不知道各位大神有何指点。
图一(右侧为marker,最亮为200bp,我加了五个孔的样品)
图二(上面的为引物二聚体)

