...expressing plasmid, such as our second most requested plasmid of all time, pMD2.G. Find psPAX2. pMD2.G -...wide range of applications. This plasmid was the top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB... top requested plasmid from 2016. You may notice the “V2.0” at the end of this plasmid which indicates...prior version of this plasmid. Indeed the first version is the 15th most popular plasmid of all time, but ...envelope expressing plasmid pMD2.G. Find pMDLg/pRPE. scramble shRNA - This plasmid from David Sabatini....1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...as they grow. The top 15 plasmids requested from Addgene psPAX2  - psPAX2 is a 2nd generation lentiviral...

...and psPAX2. Actually, these two plasmids from Didier Trono's lab are our two most popular plasmids of ...cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO plasmids you'll note that those are...influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids from Shinya... have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene ...our stride. We shipped our 100,000th plasmid. A variety of plasmids continued to be popular years after...with all the new plasmids. So far, we have distributed over 10,000 CRISPR/Cas9 plasmids to more than 3,...10 years of plasmid sharing  Interested in more facts about Addgene's 10 years of plasmid sharing? Check...

...these plasmids from Addgene. This publication then resulted in the development of a new plasmid, pCMV14...continue having access to plasmids during the COVID-19 pandemic. Over 7,000 plasmids for COVID-19 research...work to share plasmids to all scientists Resources on Addgene.org Find COVID-19 plasmids at Addgene Browse...Biological repositories that distribute mouse strains, plasmids, cell lines, and much more have been critical ...science is truly a global endeavor. Case study 2: Plasmids Another example of reagents that have broad applications...requested from Addgene. Figure 2: Total COVID-19 plasmids requested from Addgene from March 23, 2020 to ...to July 27, 2020. Browse Addgene's COVID-19 plasmids and resources! Case study 3: Mouse strains Reagents...

...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...undergoes PCR with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed...microbial contamination. Purity When possible, all plasmids used for viral production are propagated in the... system and PCR-based confirmation of transfer plasmids used in Addgene’s viral production. Barcoding ...

...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles. ...Explore our empty and EGFP retroviral plasmid set, our bacterial plasmid set for expressing tagged proteins...and AAV plasmid sets launching later this year!  Find Addgene's own Lentiviral backbone plasmids here! ... synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean, A., Chen, Y., Iyer...powerful tools in a scientist's toolkit is the empty plasmid backbone: the DNA vector that contains everything... swap sequences effortlessly.  We built these plasmids for flexibility and speed, so you can design, ...previous integrase-based approaches. Find STITCHR plasmids here! Fell, C. W., Villiger, L., Lim, J., Hiraizumi...

...integrase-deficient packaging plasmid instead of the standard packaging plasmid. For more details on how to...an integrase-deficient packaging plasmid available at Addgene: psPAX2-D64V. This construct has the D64V...lentiviral genome is distributed across multiple plasmids. For simplicity, we’ll use the second-generation...system as an example.  The lentiviral transfer plasmid encodes your insert of interest, flanked by long...this portion into the host genome. The packaging plasmid contains Gag, Pol, Rev, and Tat genes. Specifically...production of lentiviral particles.  The envelope plasmid typically encodes the vesicular stomatitis virus...into the host cell.  Transfecting these three plasmids into cells enables the production of lentiviral...

...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...scrambled shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G Envelope plasmid for producing... also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG from Robert Weinberg....1 Protocol pLKO.1 - TRC Cloning Vector Addgene Plasmid 10878. Protocol Version 1.0. December 2006. Copyright... RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine...Puromycin resistance gene for selection of pLKO.1 plasmid in mammalian cells. sin 3’LTR 3’ Self-inactivating...

... Viral Plasmids Lentiviral Plasmids Lentiviral Plasmids Additional Resources... Pooled Libraries All Viral Plasmids Packaging Plasmids Transfer Plasmids Lentiviruses, a subset of retroviruses...among multiple plasmids. Some plasmids (typically called packaging and envelope plasmids) encode components...generation transfer plasmids along with a plasmid encoding Rev Trono 12260 psPAX2 2nd Packaging Contains...conjunction with the plasmid that encodes the viral genome (typically called the transfer plasmid ). This table...packaging plasmids that can be used with 2nd and 3rd generation lentivirus technologies. ID Plasmid Generation...generation transfer plasmids Trono 12259 pMD2.G N/A Envelope VSV-G-expressing envelope plasmid Trono 8454 pCMV-VSV-G...

...Addgene CRISPR Resources CRISPR Plasmids Addgene's collection of CRISPR plasmids. CRISPR Guide Addgene's guide...sgRNA cloning lentiCRISPR v2 packaging plasmids: pVSVg , psPAX2 ; positive control: CMV-EGFP PDF, 2.4 ...lentiCas9-Blast and lentiGuide-Puro packaging plasmids: pVSVg , psPAX2 positive control: CMV-EGFP Pooled libraries...guide to using CRISPR plasmids in your lab, from knockouts to pooled library screening. CRISPR eBook Addgene's...barcoding. CRISPR Kits Addgene's collection of CRISPR plasmid kits. CRISPR Software Sanger Indel Analysis ICE... Feng Zhang Lab . Protocols Lab(s) Description Plasmids in protocol Download protocol Addgene CRISPR pooled...gRNA design and cloning for Cas9 orthologs Cas9 plasmids PDF, 107 KB Chen and Wente Zebrafish: gRNA cloning...

...DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your insert of interest...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3 pmol...pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume 500...500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be..., the plasmids should also be propagated in an endonuclease negative E. coli strain such as NEB stable...

...the transfer plasmid, a plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding adenoviral... coli . In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits. Resources...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...off-line and discarded. Confirmation of Transfer Plasmid Addgene uses a rigorous barcode matching system...microbial contamination. Purity When possible, all plasmids used for viral production are propagated in the...experiment using our virus protocols . Browse our plasmid collection of popular lentiviral vectors ....produced in 293T cells using the 2nd generation psPAX2 and pMD2.G packaging system developed by the Trono...