We narrowed to 564 results for: PAC;

...expression: pAC2 , pAC152 , pAC153 , pAC154 ; pmax dCas9 activator expression: pAC91 , pAC92 , pAC93 , pAC94... Fly: gRNA cloning pAc-sgRNA-Cas9 PDF, 169 KB Marraffini Bacteria: pCas9 new spacer cloning pCas9 PDF,...pAC94 , pAC95 ; dCas9 activator gateway donors: pAC84 , pAC1 , pAC147 , pAC148 , pAC149 ; Gateway destination...2014) (Link opens in a new window) . protoSpaceJAM protoSpaceJAM is an all-around platform for CRISPR ...destination: pAC90 PDF, 1.1 MB Katic Nematode: Cas9 and gRNA use Cas9 (pIK86) ; gRNA empty backbone (pDR274...PDF, 129 KB Marraffini Bacteria: pCRISPR new spacer cloning pCRISPR PDF, 109 KB Mendenhall and Myers Mammalian...PDF, 255 KB Zhang sgRNA cloning lentiCRISPR v2 packaging plasmids: pVSVg , psPAX2 ; positive control: CMV-EGFP...

...over to our CRISPR Plasmids and Resources page.  PAC-MAN (prophylactic antiviral CRISPR in human cells...

... (vitamin B12) Escherichia coli Remy Loris 47663 pAC-EsaR-I70V Plac 3OC6HSL (Quorum sensing molecule) ... direct recombinant proteins to the periplasmic space between the inner and outer membranes. Targeting... 113634 pSCrhaB2 PrhaB Rhamnose Burkholderia cenocepacia Miguel Valvano 47640 pCS-PesaRlux PesaR 3OC6HSL...

... Timeline Day 0: Seed 293T packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection...This procedure can be modified for alternative packaging cell lines or transfection reagents. Once produced...PES) filter Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid...Considerations Before You Start The health of the packaging cell line is critical for obtaining high viral... 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10 6 cells per plate in DMEM Complete...into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3 pmol Envelope Plasmid (...supernatant at 2100 rcf for 5 minutes to pellet any packaging cells that were collected during harvesting. Filter...

...columns have a finite binding capacity. If your sample exceeds the capacity, divide the sample among multiple...columns. For information about the total binding capacity for your columns, refer to the manufacturer’s ...Place a mark on the side of the column where the compacted resin is slanted upward. Place column in centrifuge...centrifugation step Pro-Tip Resin will appear compacted after centrifugation. Discard the flow through.... Slowly apply the sample to the center of the compact resin bed. Pro-Tip For sample volumes <1.5 mL top...the reservoir of the column. Fill the remaining space in the column with PBS and pipette several times...through into a waste container. Fill the remaining space in the column with PBS and pipette several times...

...control vector containing scrambled shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G ...particles. Note: pLKO.1 can also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG...tubes VWR: #87003-290 Note: pLKO.1 could also be packaged using pCMV-dR8.2 dvpr and pCMV-VSVG from the Robert...transfection: 1 μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl... may want to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio...

...you want to expand that with the addition of NdeI, PacI, AscI, and MfeI. With oligo overlap cloning, you...the original vector. Designing oligos To add NdeI, PacI, AscI and MfeI sites between the EcoRI and SalI ...the additional sites in tandem ( NdeI - CATATG , PacI - TTAATTAA , AscI - GGCGCGCC , MfeI - CAATTG ). ...

...Procedure The balance in the photo above has a capacity of 200 g, as indicated by the number above the...is best. Check that the balance has the correct capacity for the material that you are weighing by looking... you’re weighing out an amount larger than the capacity of your balance or weighing boat, you can weigh...

...experiment, make sure your workspace is clean and uncluttered. Disinfect your workspace with appropriate materials...times for each agent. Make sure you have enough space to work at your lab bench. It’s possible that while...

... differences, and plasmid recombination can all impact the representation of individual plasmids in the...problem as these sequences are not efficiently packaged into lentiviral particles. See below for options...overloaded. If you find that your pellet outstrips the capacity listed in the Maxiprep protocol, scale the reagent...

... appropriate for the biosafety level of the lab space Reagents Sample to be centrifuged Water Background...picture above. Wear PPE appropriate for the lab space in which you are working. Even if your samples may...

...effects. The library was cloned into the Liu lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses...be found here). To use the library, the authors packaged it into lentivirus and used the resulting lentiviral...large size of dCas9-VPR limits one’s ability to package it into lentivirus and later create stable cell...cells. The piggybac system has a much larger cargo capacity than lentivirus and is capable of efficiently ...

..., viral packaging eligibility, and more. ID Plasmid Description PI Return to top AAV Packaging Plasmids...encoded in the packaging plasmid will determine the AAV vector serotype. Some packaging plasmids also ...the helper plasmid. Packaging Plasmids Available with Addgene's Viral Vector Packaging Service This table...table lists the packaging plasmids (Rep/Cap plasmids) used by Addgene's Viral Vector Packaging Service to ...AAV packaging plasmid, expressing Rep2 and Cap1 James M. Wilson 232199 pAAV2/2 AAV2 AAV packaging plasmid...AAV packaging plasmid, expressing Rep2 and Cap7 James M. Wilson 112864 pAAV2/8 AAV8 AAV packaging plasmid...Gradinaru All Available AAV Packaging Plasmids This table lists all packaging plasmids (Rep/Cap plasmids...

...Huiwang Ai 73544 pEvol-pAcFRS.2.t1 pAcFRS.2.t1 E. coli p-acetyl-l-phenylalanine (pAcF) Bacterial TAG Farren... Isaacs 73545 pEvol-pAcFRS.1.t1 pAcFRS.1.t1 E. coli p-acetyl-l-phenylalanine (pAcF) Bacterial TAG Farren...141173 pAcBac1-3nitroY-A7-RS A7 3nY tRNA synthetase M. barkeri Mammalian Ryan Mehl 141174 pAcBac1-haloTyrRS...acid Bacerial, Mammalian TAG Peter Schultz 50831 pAcBac2.tR4-OMeYRS/GFP* tyrosyl-tRNA synthetase E. coli... amino acids Mammalian TAG Peter Schultz 50832 pAcBac1.tR4-MbPyl PylRS M. barkeri variety of unnatural...barkeri Tetrazine 3.0 Bacterial TAG Ryan Mehl 174081 pAcBac1-NES-Flag-R2-84-MbRS Tetrazine3.0 NES-Flag-R2-84...barkeri acridone Bacterial TAG Ryan Mehl 174101 pAcBac1.tR4-AcdRS82 Acridone 82 tRNA synthetase M. barkeri...

... transgene, packaging capacity, and size. Vector Type Expression Genome Packaging Capacity Virus Size ...host genome. AAVs are limited by their small packaging capacity. Gamma-retroviral Vector Guide | Gamma-retroviral...Adenoviral Plasmids Adenoviral vectors have high packaging capacity, high transduction efficiency, and high but...Interested in Addgene's viral services? See Addgene's Packaged on Request service for made-to-order AAV vectors...engineered such that they can be delivered to packaging cell lines and use their cellular machinery to...the viral genome to assemble the viral capsids (packaging and envelope plasmids). When transfected into ...Resource Guide . Resources Addgene’s Viral Vector Packaging Service Addgene's Viral Protocols Addgene Blog...

...referred to as CRISPR ( C lustered R egularly I nterspaced S hort P alindromic R epeat) technologies. Before...for Cas-binding and a user-defined ∼20-nucleotide spacer that defines the genomic target to be modified....target is present immediately adjacent to a P rotospacer A djacent M otif (PAM) The PAM sequence (NGG)... an active, DNA-binding configuration, with the spacer region of the gRNA left free to interact with the...Cas9 will only cleave a given locus if the gRNA spacer sequence shares sufficient homology with the target...throughout the genome, called off-targets, that can impact your experiment. There are many online tools available...Cas9’s interactions with DNA phosphate backbone HypaCas9 — increase Cas9 proofreading and discrimination...

...shRNA of interest flanked by LTRs Packaging plasmid — contains packaging and regulatory genes; one plasmid...Addgene lentiviral packaging, envelope, and transfer plasmids Viral...Lentiviral Plasmids Transfer Plasmids Packaging Plasmids Envelope Plasmids Resources Lentiviruses, a subset... third-generation Envelope plasmid — contains packaging gene env This collection highlights popular second...second- and third-generation lentiviral packaging, envelope, and transfer plasmids available at Addgene.... Feng Zhang Return to top Popular Lentiviral Packaging Plasmids ID Plasmid Generation Description PI ...generation transfer plasmids along with additional packaging components. Didier Trono 12260 psPAX2 2nd Contains...

...contains many unique protospacer sequences that have homology to foreign DNA. Protospacers are separated by...Adaptive Immune System CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) sequences were initially...contain a species-specific sequence known as a protospacer adjacent motif (PAM). The CRISPR complex binds...sequence is only valid if it contains a special Protospacer Adjacent Motif (PAM) directly after where the... development has occurred at a truly astonishing pace, with work directed towards increasing on-target...fidelity Cas9 variants , SpCas9-HF, eSpCas9, and HypaCas9 display very low off-target activity due to rationally...SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9 orthologs also have distinct PAM ...

...remain in the laboratory. When traveling between lab spaces, take your glove off of one hand and use your bare...

...at a lower voltage. You will want to have enough space around each band to cut without having DNA in other...