Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 26966 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV1 | 26966-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV5 | 26966-AAV5 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information | Add to Cart | |||
| AAV9 | 26966-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)
- Backbone sizew/o insert(bp)5609
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)NEB Stable
- Growth instructionsPlease use Rec A- competent cells such as Stbl3 cells from Invitrogen for transformation.
- Copy numberHigh Copy
Gene/Insert
- Gene/Insert nameeNpHR 3.0
- SpeciesH. sapiens (human)
- Insert Size (bp)1683
- MutationER export at 3' end of gene. Trafficking signal (TS) in between gene and fluorophore.
- Tag/ Fusion Protein
- EYFP (C terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteAscI(not destroyed)
- 3′ cloning siteNheI(not destroyed)
- 5′ sequencing primerggccagcttggcacttgatg (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
- Articles Citing this Plasmid
- 14 References
Depositor Comments
This plasmid contains the human elongation factor-1a promoter. Golgi export trafficking signal (TS) sequence is KSRITSEGEYIPLDQIDINV. ER export sequence is FCYENEV.
For additional information please visit - http://www.optogenetics.org
Information for AAV1 (Catalog # 26966-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV-Ef1a-DIO eNpHR 3.0-EYFP (#26966). In addition to the viral particles, you will also receive purified pAAV-Ef1a-DIO eNpHR 3.0-EYFP plasmid DNA.
EF1a-driven, cre-dependent eNpHR 3.0-EYFP for optogenetic inhibition.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 GeneEYFP (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.
Information for AAV5 (Catalog # 26966-AAV5)(Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV-Ef1a-DIO eNpHR 3.0-EYFP (#26966). In addition to the viral particles, you will also receive purified pAAV-Ef1a-DIO eNpHR 3.0-EYFP plasmid DNA.
EF1a-driven, cre-dependent eNpHR 3.0-EYFP for optogenetic inhibition.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, AAV5 cap gene
- BufferPBS + 0.001% Pluronic F-68
- SerotypeAAV5
- PurificationIodixanol gradient ultracentrifugation
- Reporter GeneEYFP (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
- 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.
Information for AAV9 (Catalog # 26966-AAV9)(Back to top)
Purpose
Ready-to-use AAV9 particles produced from pAAV-Ef1a-DIO eNpHR 3.0-EYFP (#26966). In addition to the viral particles, you will also receive purified pAAV-Ef1a-DIO eNpHR 3.0-EYFP plasmid DNA.
EF1a-driven, cre-dependent eNpHR 3.0-EYFP for optogenetic inhibition.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
- Reporter GeneEYFP (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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便携式pH计主要用于现场和野外测方式，要求较高的精度和完善的功能。
实验室pH计是一种台式高精度分析仪表，要求精度高、功能全，包括打印输出、数据处理等等。
工业pH计是用于工业流程的连续测量，不仅要有测量显示功能，还要有报警和控制功能，以及安 装、清洗、抗干扰等等问题的考虑。
2.据ph计仪器精度分类：可分为0.2级、0.1级、0.05级、0.01级,数字越小，精度越高。
3.根据元器件类型分类：可分为晶体管式、集成电路式和单片机微电脑式，现在更多的是应用微电脑芯片，大大减少了仪器体积和单机成本；但芯片的开发成本很贵。
4根据读数指示分类：可分为指针式和数字显示式二种。指针式pH计现在已很少使用，但指针式仪表能够显示数据的连续变化过程，因此在滴定分析中还有使用。
5.看PH计有没有附带功能，比方说带标配RS232接口，还有一个很重的是温度补偿是自动
还是手动，自动温度补偿的PH计要比手动温度补偿的PH计要方便些，二者之间的价格你就明白了吧。
斜率的计算是和电极测量你所配制的溶液的电位有关系的。
三个标准缓冲液，标定的时候一般选择两个就行，如果三个都参与标定，斜率会相应的有差别。
电极的话，分立的电极是参比和测量各一支，分别接在仪器后面，使用时一起测量。复合电极，有的是参比和测量复合电极，应该只有一个球泡。还有一种三复合电极，是参比，测量，还有温度都集于一支电极上，上面除了玻璃球泡，那个小黑的是测量溶液温度的。
进口品牌JENCO的性价比比较高。
按测量精度
可分0.2级、0.1级、0.01级或更高精度。
按仪器体积
分为笔式（迷你型）、便携式、台式还有在线连续监控测量的在线式。
根据使用的要求
笔式（迷你型）与便携式pH酸碱度计一般是检测人员带到现场检测使用。
选择pH酸碱度计的精度级别是根据用户测量所需的精度决定，而后根据用户方便使用而选择各式形状的pH计。
◆按便携性分的，分为：便携式pH计，台式pH计和笔式pH计。
◆按用途分为：实验室用pH计，工业在线pH计等。
◆按先进程度分为经济型pH计，智能型pH计，精密型pH计或分为指针式pH计，数显式pH计。
◆笔式pH计，一般制成单一量程，测量范围狭，为专用简便仪器。
便携式和台式pH计测量范围较广，常用仪器，不同点是便携式采用直流供电，可携带到现场。实验室pH计测量范围广、功能多、测量精度高。
工业用pH计的特点是要求稳定性好、工作可靠，有一定的测量精度、环境适应能力强、抗干扰能力强，具有模拟里量输出、数字通讯、上下限报警和控制功能等。向左转|向右转
玻璃电极在初次使用前，必须在蒸馏水中浸泡一昼夜以上。
平时也应浸泡在蒸馏水中以备随时使用。玻璃电极不要与强吸水溶剂接触太久，在强碱溶液中使用应尽快操作，用毕立即用水洗净，玻璃电极球泡膜很薄，不能与玻璃杯及硬物相碰；玻璃膜沾上油污时，应先用酒精，再用四氯化碳或乙醚，最后用酒精浸泡，再用蒸馏水洗净。注意电极内要充满氯化钾溶液，应无气泡，防止断路。应有少许氯化钾结晶存在，以使溶液保持饱和状态，使用时拨去电极上顶端的橡皮塞，从毛细管中流出少量的氯化钾溶液，使测定结果可靠。
另外，pH测定的准确性取决于标准缓冲液的准确性。酸度计用的标准缓冲液，要求有较大的稳定性，较小的温度依赖性。
测定时把复合电极插在被测溶液中，由于被测溶液的酸度（氢离子浓度）不同而产生不同的电动势，将它通过直流放大器放大，最后由读数指示器（电压表）指出被测溶液的pH值。酸度计能在0～14pH值范围内使用。酸度计有台式、便携式、表型式等多种，读数指示器有数字式和指针式两种.
1、校正：先将仪器斜率调节器调节在100％位置，再根据被测溶液的温度，调节温度调节器到该温度值。
2、定位：把复合电极插入仪器，选择一种最接近样品pH值的缓冲溶液，把电极放入这一缓冲溶液里，摇动烧杯，使溶液均匀.待读数稳定后，该读数应是缓冲溶液的pH值，否则就要调节定位调节器。用于分析精度要求较高的测定时，要选择两种缓冲溶液（即被测样品的pH值在该两种缓冲溶液的pH值之间或接近）。待第种缓冲溶液的pH值读数稳定后，该读数应为该缓冲溶液的pH值，否则调节定位调节器。清洗电极，吸干电极球泡表面的余液。把电极放入第二种缓冲溶液中，摇动烧杯使溶液均匀，待读数稳定后，该读数应是第二种缓冲溶液的pH值，否则调节斜率调节器。
3、测量：经过pH标定的仪器，即可用来测定样品的pH值。这时温度调节器、定位调节器、斜率调节器都不能再动，用蒸馏水清洗电极，用滤纸吸干电极球部后。把电极插在盛有被测样品的烧杯内，轻轻摇动烧杯，待读数稳定后，就显示被测样品的pH值。复合电极的主要传感部分是电极的球泡，球泡极薄，千万不能跟硬物接触。测量完毕套上保护帽，帽内放少量补充液（3mol/L的氯化钾溶液），保持电极球泡湿润.仪器采用CMOS集成电路，不用时插入短路插头。检修时电烙铁要有良好的接地，以保护仪器。
