Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Plasmid | 100896 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $75 | Add to Cart | |
| AAV5 | 100896-AAV5 | Limited Stock Available, 4 units leftVirus (100 µL 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(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 nameEGFP
- PromoterGFA104 (GFAP promoter)
Cloning Information
- Cloning methodUnknown
- 5′ sequencing primerunknown (Common Sequencing Primers)
Resource Information
- Supplemental Documents
- pAAV.GFA104.PI.eGFP.WPRE.bGH (p2380)-Q.gb
- Terms and Licenses
- UBMTA
- Ancillary Agreement for Plasmids Containing FP Materials
- Industry Terms
- Not Available to Industry
Depositor Comments
This plasmid is also known as pAAV.GFA104.PI.eGFP.WPRE.bGH (p2380) from the Penn Vector Core.
Information for AAV5 (Catalog # 100896-AAV5)(Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV.GFA104.PI.eGFP.WPRE.bGH (#100896). In addition to the viral particles, you will also receive purified pAAV.GFA104.PI.eGFP.WPRE.bGH plasmid DNA.
GFA104-driven eGFP expression. 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
- Reporter GeneEGFP
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.
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1、测浓度和纯度,是否达标
2、跑电泳看条带是否与测的数据相互印证
如果上述两点满足,就不需要纯化,否则需要进一步纯化。
不过根据经验,一般都不需要,我们实验室用的是GNT系列的试剂盒,提取结果就直接进入下游实验了,效果挺稳定
质粒是真核细胞细胞核外或原核生物拟核区外能够进行自主复制的遗传单位,包括真核生物的细胞器(主要指线粒体和叶绿体)中和细菌细胞拟核区以外的环状脱氧核糖核酸(DNA)分子.现在习惯上用来专指细菌(大肠杆菌)、酵母菌和放线菌等生物中细胞核或拟核中的DNA以外的DNA分子.
小鼠没有叶绿体,你可以直接参照小鼠线粒体的提取方法进行提取.
样品处理:
a. 植物组织:取新鲜或-70℃冻存100mg组织在液氮中研磨,把粉末加入到1ml裂解液中混匀。
b. 动物组织:取新鲜或-70℃冻存100mg组织加1ml裂解液,用组织研磨杵或匀浆器匀浆处理。
c. 贴壁细胞:直接在培养板中加入裂解液裂解细胞,每106细胞加1ml 裂解液。用取样器吹打混匀。
人教版高中生物必修二有这个图
植物材料 用最精确的方法,称取不超过100mg的植物材料,置于处理过的研钵,加入液氮进行研磨 将研磨得到的粉末,快速转移至无RNase,并经过液氮冷却的2mL离心管(自备),
目的掌握RNA提取的基本技术,了解RNA提取过程中的各种注意事项。二、原理RNA提取是分子生物学实验中难度较大的实验技术。RNA提取和DNA提取有类似的地方,因为它们都是核酸,都具有较好的水溶性。提取RNA首先破碎细胞,然后用提取液将RNA溶出,反复抽提去除蛋白质 ……

