We narrowed to 563 results for: pac
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TypeBlog Post... important to have nice crisp bands and to have space to cut out the bands. Because of this we recommend...
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Finding Your Perfect Job After University
TypeBlog Post...I really loved the fact that my work had a real impact people’s lives and would improve their treatment... -
RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog Post...the most prevalent method due to ease of use and capacity to generate a long-term RNAi phenotype. Long hairpin... -
Antibodies 101: Validation
TypeBlog Post...specificity. However, there are countless variables that impact an antibody’s affinity and specificity for a given... -
Deep Dive: Statistical Tests (Comparisons)
TypeBlog Post...independent variables. For example, you could test the impact of temperature, humidity, sunlight exposure time... -
Which Fluorescence Microscopy Technique is Best for Me?
TypeBlog Post...imaging, the light microscope was limited in its capacity to resolve closely lying structures. This is because... -
Custom CRISPR Screens & the Green Listed Software
TypeBlog Post...GC%, reverse compliment sequences, and more. Output_Compact: A text file that describes which genes were... -
Antibodies 101: Introduction to Antibodies
TypeBlog Post...stability. Additionally, the choice of vector can impact the expression level of an antibody (Ayyar et al... -
Brain Initiative Collection
TypeCollection...65417-AAV8 pAAV-hSyn-dF-HA-KORD-IRES-mCitrine AAV packaging vector for hSyn driven KOR DREADD expression. ... -
A Needle in a Base-Stack: Cas9 Structural Biology
TypeBlog Post...important to choose a target sequence with a nearby protospacer-adjacent motif (PAM)? Cas9 is becoming an ever... -
How to Deposit Your Plasmids with Addgene
TypeBlog Post...with a unique Deposit ID. Addgene will send you a package containing instructions on how to prepare your ... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post...optimal scaffold can depend on the length of the spacer region between the target sequences. These updates... -
Lentiviral Vector Guide
TypeGuide...flanked by LTRs; 8–10 kb packaging capacity Packaging plasmid — contains packaging genes gag and pol , and...tat gene Can be packaged by both second- and third-generation packaging systems Packaging Plasmid One plasmid...isoforms of these packaging components. Therefore, lentivirus may not be efficiently packaged by gamma-retroviral...transgene and wild-type LTRs Packaging plasmid — contains entire viral genome (packaging, regulatory, and accessory...and LTRs (chimeric 5’ LTR) Packaging plasmid 1 — contains gag and pol Packaging plasmid 2 — contains rev...transfer plasmids can be packaged by either second- or third-generation packaging plasmids. For a summarized...Third-generation Transfer Plasmid Can be packaged ONLY by a second-generation packaging system that includes the tat... -
Gamma-Retroviral Vector Guide
TypeGuide...flanked by LTRs; ~8 kb packaging capacity Packaging plasmid — contains packaging genes gag and pol Envelope...of these packaging components. Therefore, gamma-retroviruses may not be efficiently packaged by lentiviral...to lentiviral packaging methods . The three plasmids described above (envelope, packaging, and transfer... . Production Using Packaging Cell Lines Gamma-retroviral vectors can be packaged using helper-free cell...envelope and packaging plasmids. Unless multiple recombination events occur between the packaging, envelope...simpler genomes, containing only the necessary packaging genes, while lentiviruses also contain accessory...lentiviruses and gamma-retroviruses use the same packaging genes. However, they are different viruses and... -
Adenovirus Guide
TypeGuide...different plasmids or the packaging cell line, which not only frees up more space for genetic cargo but also...vectors and further increase the transgene packaging capacity up to ~10.5 kb. The deleted E2 and E4 regions...linearized with PacI to create a linear dsDNA construct flanked by ITRs. Cells from the packaging HEK293 or ...contain the viral gene E4, adding 2.7 kb of packaging space. However, these constructs must be transfected ...rAdV vectors can provide a large transgene packaging capacity. Large protein-coding genes and complex gene...guide RNA and other regulatory elements. The packaging capacity of rAdV vectors (~6.5 kb for first-generation...included between the ITRs is limited by the physical space available inside of the adenoviral capsid and directly... -
Adeno-associated virus (AAV) Guide
TypeGuide...cell DNA synthesis but further limits the packaging capacity of rAAV vectors to ~2.4 kb. Figure 4: Examples... capsid is challenging due to its limited packaging capacity, especially if any additional regulatory ... more information. Split AAVs to increase packaging capacity Another strategy to deliver cargo too large...and References Addgene's Viral Vector Packaging Services Packaged-on-Request AAV Preps In-Stock AAV Preps...& Engelhardt, J. F. (2001). Expanding AAV packaging capacity with trans-splicing or overlapping vectors... are involved in genome replication and viral packaging. The Cap region encodes three structural capsid...origins of replication and also act as signals for packaging. Figure 1: Wild-type AAV genome. Created with ... -
CRISPR Guide
TypeGuide...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... -
Plan Your Experiment
TypeGuide...vectors is that the packaging limit (how large of a DNA sequence you can actually package into your viral ...Edit References CRISPR ( C lustered R egularly I nterspaced S hort P alindromic R epeats) is a powerful system...efficiencies. The template sequences can also have a large impact on editing efficiency. So while testing multiple...promoters, and selection markers. There are also no packaging limits as there are for viral vectors — the only... Addgene's in-stock AAV preps or try Addgene's Packaged on Request service. Delivery Using mRNA and Proteins... -
Sequencing Primers
TypeGuide...pBABE 5' CTTTATCCAGCCCTCAC (Weinberg Lab) Psi packaging signal, 5' of MCS in pBABE vectors, forward primer...CGCAACGATCTGGTAAACAC (Invitrogen) OpIE2 promoter, forward primer pACYC-F TGAAGTCAGCCCCATACGAT p15A origin, forward primer... -
Antibody Guide
TypeGuide...antibodies are collected, tested against the antigen, packaged, and sold. This is the most cost-efficient way...antibodies to upwards of twenty. Laser and software capacity theoretically allow for roughly fifty different...