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Item | Catalog # | Description | Quantity | Price (USD) | ||
---|---|---|---|---|---|---|
Plasmid | 51084 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
AAV1 | 51084-AAV1 | Virus (100 µL at titer ≥ 5×10¹² vg/mL)and Plasmid.More Information | Add to Cart | |||
AAV Retrograde | 51084-AAVrg | Virus (100µL at titer ≥ 7×10¹² vg/mL)and Plasmid.More Information | Add to Cart |
This material is available to academics and nonprofits only.
Backbone
- Vector backbonepAAV-hSyn1-WPRE-HGHpA(Search Vector Database)
- Backbone manufacturerModified from Karl Deisseroth
- Backbone sizew/o insert(bp)4590
- Total vector size (bp)6750
- Modifications to backboneAltered cloning sites
- Vector typeMammalian Expression, AAV
Growth in Bacteria
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)Stbl3
- Copy numberHigh Copy
Gene/Insert
- Gene/Insert nameGCaMP6s
- Alt nameG6s
- Alt nameGCaMP6 slow
- SpeciesSynthetic
- Promoterhuman Synapsin1
- Tag/ Fusion Protein
- P2A-nls-dTomato (C terminal on insert)
Cloning Information
- Cloning methodRestriction Enzyme
- 5′ cloning siteBamHI(not destroyed)
- 3′ cloning siteAscI(not destroyed)
- 5′ sequencing primerctcagcgctgcctcagtctg
- 3′ sequencing primerCCATACGGGAAGCAATAGCATG (Common Sequencing Primers)
Resource Information
- A portion of this plasmid was derived from a plasmid made byThe GCaMP6s portion was derived from Addgene plasmid #40753 (Douglas Kim, Janelia Farms).
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- Takara Bio Limited Use Label License (formerly Clontech)
- Industry Terms
- Not Available to Industry
- Articles Citing this Plasmid
- 2 References
Depositor Comments
The addition of the P2A-nls-dTomato allows for easy identification of GCaMP6 expressing cells exhibiting nuclear red fluorescence that does not significantly overlap with the cytosolic GCaMP signal.The GCaMP and fluorophore are physically uncoupled.
Permissions were obtained from Douglas Kim and the Clontech licensing office for depositing this new plasmid.
Information for AAV1 (Catalog # 51084-AAV1)(Back to top)
Purpose
Ready-to-use AAV1 particles produced from AAV-hSyn1-GCaMP6s-P2A-nls-dTomato (#51084). In addition to the viral particles, you will also receive purified AAV-hSyn1-GCaMP6s-P2A-nls-dTomato plasmid DNA.
GCaMP6s calcium sensor and bicistronic, physically separate nuclear localized dTomato expression under a human synapsin1 promoter. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume100 µL
- Titer≥ 5×10¹² 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, AAV1 cap gene
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV1
- PurificationIodixanol gradient ultracentrifugation
- Reporter Genenuclear dTomato (physically separate, not a fusion protein)
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
Data submitted about 51084-AAV1 by requesting scientist(s):
- Data 1: Human (ex: human brain slices), Infected cells in vitro
Information for AAV Retrograde (Catalog # 51084-AAVrg)(Back to top)
Purpose
Ready-to-use AAV Retrograde particles produced from AAV-hSyn1-GCaMP6s-P2A-nls-dTomato (#51084). In addition to the viral particles, you will also receive purified AAV-hSyn1-GCaMP6s-P2A-nls-dTomato plasmid DNA.
GCaMP6s calcium sensor and bicistronic, physically separate nuclear localized dTomato expression under a human synapsin1 promoter. These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons.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, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
- BufferPBS + 0.001% Pluronic F-68 + 200 mM NaCl
- SerotypeAAV retrograde (AAVrg)
- PurificationIodixanol gradient ultracentrifugation
- Reporter Genenuclear dTomato (physically separate, not a fusion protein)
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
Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.Data submitted about 51084-AAVrg by requesting scientist(s):
- Data 1: Rat, Brain parenchyma
- Data 2: Mouse, Brain parenchyma
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1.我利用氯仿萃取生物碱,需要将氯仿蒸干,我做的样品有非常多份,但是每份样品很少,用的氯仿也很少,每份只用十几毫升左右,但是旋转蒸发仪的蒸发瓶好大,而且一次只能蒸发一个样,太慢了,就算蒸发完了,取出生物碱的时候也不好取,由于我做的是定量试验,所以如果取出生物碱时有残留会影响实验结果,到底怎么办啊?有没有特制的小的蒸发瓶?或者特制的可以一次接好几个蒸发容器的接口?
2.请问恒温水浴锅可不可以用来做蒸发?由于是蒸发氯仿,担心没有回收装置会导致泄露中毒的问题,但是我的量比较少,又想冒险试试,真是焦头烂额了现在,大家把那个我出出主意吧!!!!
一背景:最近正用水饱和正丁醇萃取某海洋生物乙醇提取物,考虑活性问题,温度一般不超过56度。
二问题:萃取液在开始蒸发效果还好,但超过一个钟后慢下来,最后几乎不滴。很多高手提到再40度就可以蒸发,不到五十度就可以蒸去大部。我们定在五十几度也一样,慢慢发现停机休息几个小时又可以蒸发了,但过段时间又慢下来,百思不得解。
三折腾:于是开始试验水饱和正丁醇的制备是否有问题、蒸发效果跟时间及水浴温度的关系、蒸发剩余加水多少。。。。毫无进展,郁闷,有时这机器休息时间短些还不行,真想烧香拜佛了。
四辛勤加运气:一日,感觉水泵水箱水温有点高(没有象汽车那样弄水循环,一般干些就加满),担心电动机烧坏于是换凉水,突然发现冷凝管液滴增多,下滴速度加快了,终于找到原因:水泵抽吸出问题了!可能是工作时间长了,水泵电动机发热,如果温度升高,抽吸效果下降。虽然仪器指示负压很高了,其实没有起到很大作用,比如关掉水泵,只要不开气阀负压也不会降下来的。
五解决:马上使用循环水降低水箱温度。问题解决,但新的问题来了:进出水速度一致难调而且靠近水箱上缘、感觉效果不好。素性将冷凝水直接加入水箱,又省水又达到目的:可是水的多少不好控制。解决:直接把水泵水箱置于水池中(支架上)。
五思考:1实验中首先要对仪器的使用性能有所了解。后仔细观察,其实此水泵是可以进出水的(循环),只是按一般的使用习惯没有使用循环水,而只是动手添加水。奉劝各实验室水泵改循环水,特别是需要较长时间运作时,(形式多样,保证水泵长时间工作就好),而不是添加水,否则,使用时间长了会跟我们一样郁闷折腾。
2现象的存在总是有原因、可以解释的。多观察、多动手、多对比各种差异有助于找到某种原因。
3不要过分相信仪器显示的数据:起码这回我们被骗了:负压显示那么高,一直不怀疑水泵抽吸的效果;从这启发:另一同志仔细测水浴锅的温度,发现居然相差二、三度左右!二、三度对提取海洋生物药来讲简直是“差之毫厘,谬以千里”!
最后多谢捧场!呵呵
我用乙醇来提取,但是回收瓶中只有乙醇,那里出问题了呢????
祝大家元宵节快乐!
已经试过的方法有:电吹风烘箱
将试的方法是:超声
大家有什么好建议,谢谢了!
我们有个中药项目,需要采用乙酸乙酯萃取,有效成分分布在乙酸乙酯有机层中,现在我们采用旋转蒸发仪回收乙酸乙酯直到蒸发瓶中有效成分成蜂窝固体状才取出,但现在有一个问题,我如何判断蒸发瓶中蜂窝状固体可以刮出?目前我们实验室做的时候是根据蒸发瓶释放真空时蜂窝状固体不收缩的标准作为蒸干取出固体物的,但这个标准太粗放了,我也知道光靠旋转蒸发仪是不可能除尽有机溶剂,我们后续还有一个工艺可以除去蒸干固体残留的有机溶剂。我现在想知道有没有一些判断标准用于旋转蒸发仪固体蒸干可以取出来的标准可供参考啊?盼指教,谢谢
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