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We narrowed to 13 results for: plasmid dna

Showing: 1 - 13 of 13 results
  1. Molecular Biology Reference

    Type
    Guide
    ...propagating plasmids Antibiotics commonly used for plasmid selection DNA sequencing for plasmid verification...fragment of DNA to be studied into the plasmid. Below are some common plasmid elements: Plasmid Element Description...retain the plasmid DNA, despite the added replication burden, as bacteria without the plasmid DNA would not...Restriction Cloning Plasmids 101: Inducible Promoters Plasmids 101: What is a plasmid? Plasmids 101: Antibiotic...Molecular Genetics Plasmid Elements Working with Plasmids Introduction Types of Plasmids E. coli strains ...host cell. Like the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division, and each daughter...recombinant DNA, in which many identical DNA fragments can be generated. The combination of a DNA fragment...
  2. Cloning

    Type
    Guide
    ...ends in the DNA fragments. Given its prevalence, the vast majority of plasmids used for DNA cloning and... your DNA fragment of interest. Now that your DNA fragment has been cloned into a donor plasmid, it can...nucleotides and lastly DNA ligase fuses DNA strands together. In Gibson assembly DNA fragments with 20-40...archaea, cleave double stranded DNA (dsDNA) at specific sequence sites in the DNA. In restriction cloning, scientists...end of dsDNA fragments to generate 3' single-stranded overhangs. DNA polymerases synthesize DNA molecules...them into plasmids for replication and protein expression. So how do scientists recombine DNA? There are...any piece of DNA that already has restriction sites on either side of it into any plasmid that has the...
  3. CRISPR Guide

    Type
    Guide
    ...purify specific regions of DNA, image DNA in live cells, and precisely edit DNA and RNA. The ease of generating...Browse Plasmids: RNA Targeting , RNA Editing Degrading DNA with Cas3 Rather than initiating DNA cleavage...-stranded DNA plasmid. Small ssDNA oligos often work well for small edits like a single-point mutation...homology with the target DNA. Once the Cas9-gRNA complex binds a putative DNA target, the seed sequence...will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...strands of the target DNA. This results in a double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides...of target DNA. Cas9n increases target specificity. Since two nickases targeting opposite DNA strands are...
  4. Promoters

    Type
    Guide
    ...biology basics and plasmid components. Plasmids 101: The Promoter Region-Let's Go Plasmids 101: Inducible ...the binding of RNA polymerase to DNA. RNA polymerase transcribes DNA to mRNA which is ultimately translated...that the DNA tends to fold back on itself which helps to explain how many physically distant DNA sequences... popular Plasmids 101 series , covers topics ranging from the newest breakthroughs in plasmid technologies...Inducible Promoters Plasmids 101: Repressible Promoters Plasmids 101: Terminators and PolyA signals... Plasmid Reference Molecular Biology Reference Promoters...Promoters Promoters Definition A promoter is a region of DNA where transcription of a gene is initiated. Promoters...
  5. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...different plasmid. In order to produce rAAV vectors, three plasmids are needed: Transfer plasmid (also known...the packaging plasmid. For a summary of all AAV plasmid components, see the AAV Plasmid Elements table...repeat DNA molecule that spontaneously reanneals upon infection, forming a double-stranded DNA (dsDNA) template...-stranded DNA genome. Adeno-associated viruses (AAV), however, are small single-stranded DNA viruses that...as cis plasmid) — containing the transgene of interest between the two ITRs. Packaging plasmid (also known...chosen plasmid is compatible with cloning methods available in your lab. When cloning your plasmids, be ... often results in a greater yield of DNA. However, if the plasmid contains a Gateway cassette containing...
  6. Adenovirus Guide

    Type
    Guide
    ...you generally need two plasmids: Shuttle plasmid (also known as transfer plasmid) — containing the transgene...shuttle plasmid. For a summary of all adenoviral plasmid components, see the Adenoviral Plasmid Elements...for viral DNA replication and host cell manipulation. Late transcripts are expressed after DNA replication...-stranded DNA genome. Adeno-associated viruses (AAV), however, are small single-stranded DNA viruses that...for recombination with the adenoviral plasmid. Adenoviral plasmid — containing the rest of the adenoviral...chosen plasmid is compatible with cloning methods available in your lab. When cloning your plasmids, be ... often results in a greater yield of DNA. However, if the plasmid contains a Gateway cassette containing...
  7. Plan Your Experiment

    Type
    Guide
    ...design. Addgene offers many plasmids containing validated gRNAs . These plasmids contain gRNAs that have ...generate stable cell lines if your plasmid contains a selection marker. Plasmid delivery is versatile thanks...more than just the DNA sequence. If this is the case, you can confirm your edit using DNA-based methods like...Application Cas Enzyme Additional Considerations Browse Plasmids Knockout Permanently disrupt gene function; cells...mutations or small tag insertions, by copying from a DNA donor template Cas9 or Cas9 fusions with proteins...different activator systems exist, including the multi-plasmid SAM system Activate Design and Clone Your gRNA ...efficiency . Remember that HDR also requires donor DNA to serve as template for the repair. Once you settle...
  8. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...Gamma-Retroviral Plasmid Elements Plasmid Type Element Delivery relative to transgene Purpose Transfer plasmid LTR ...transcribed into DNA before being integrated in the genome of a host. Going from RNA to DNA is the reverse... of all gamma-retroviral plasmid components, see the Gamma-Retroviral Plasmid Elements table . In order...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or ...capacity Packaging plasmid — contains packaging genes gag and pol Envelope plasmid — contains packaging...chosen plasmid is compatible with cloning methods available in your lab. When cloning your plasmids, be ... often results in a greater yield of DNA. However, if the plasmid contains a Gateway cassette containing...
  9. Lentiviral Vector Guide

    Type
    Guide
    ...second-generation transfer plasmids as well. Third-generation plasmids include (Figure 4): Transfer plasmid — contains...Packaging plasmid 1 — contains gag and pol Packaging plasmid 2 — contains rev Envelope plasmid — contains...systems Packaging Plasmid One plasmid encoding gag , pol , tat , and rev Two plasmids, one encoding gag...Lentiviral Plasmid Elements Plasmid Type Element Delivery relative to transgene Purpose Transfer plasmid LTR ...transcribed into DNA before being integrated in the genome of a host. Going from RNA to DNA is the reverse...separated into distinct plasmids for safety. For a summary of all lentiviral plasmid components, see the ...four, for third generation) plasmids: Transfer plasmid — contains transgene, sgRNA, or shRNA of interest ...
  10. Guide to Using Pooled Libraries

    Type
    Guide
    ... In cDNA libraries, each plasmid contains a unique cDNA. In shRNA or gRNA libraries, each plasmid contains... Science Guides Guide to Plasmid Pooled Libraries Plasmid Pooled Libraries Jump To: Pooled Library...are single preparations of many different plasmids. Plasmids within a given library have the same backbone...protocol for the best results. For some libraries, plasmid DNA can be delivered directly to the cells of interest... others, notably pooled lentiviral plasmid libraries, the plasmids must first be used to make virus. This...infected cell receives only one plasmid. To make sure that every plasmid is adequately represented in the... given plasmid, think of each cell carrying it as being a biological replicate for that plasmid. Library...
  11. Antibody Guide

    Type
    Guide
    ...little variation. Recombinant plasmids - Antibodies can be produced from plasmids, with an antibody’s genetic... genes from specific hybridoma cells into plasmids. Plasmids are then transfected into cells and induced...interactions between DNA and protein. Micrococcal nuclease digestion is used to shear the DNA in these assays... uses antibodies to capture protein:DNA complexes. After capture, the DNA is released from the complex...suitable for DNA:histone modifier interactions, which have strong binding between the protein and DNA. Some ...immune cells, or through the use of recombinant plasmids. Animals - Antibodies produced in animals make...genetic code inserted into the plasmid. This produces the least amount of variation between antibodies, allows...
  12. Sequencing Primers

    Type
    Guide
    ... For sequencing plasmids in our repository, we've chosen primers based on the plasmid backbone and insert...uses for sequence verification of deposited plasmids. Plasmid... Plasmid Reference Molecular Biology Reference Sequencing... useful in your sequencing reaction, find your plasmid page and see what primers are listed under "5' ... for sanger sequence verification of deposited plasmids. Below is a list of commonly used primers. This...primer Gal4 N-term GAGTAGTAACAAAGGTCAA 3' end of Gal4 DNA binding domain, forward primer Gal4-AD AATACCACTACAATGGAT...CGTCAGCAGAGCTTCACCATTG (Caldwell lab) 3' end of LexA DNA binding domain, forward primer LKO.1 5' GACTATCATATGCTTACCGT...
  13. Optogenetics Guide

    Type
    Guide
    ... check out our Biosensors Plasmid Collection . To find optogenetics plasmids available through Addgene...mutagenesis - use Addgene's Optical Switch Plasmid Table to search for plasmids for your next experiment. For a ...domain (iLID plasmid), while its natural binding partner SspB is included in a separate plasmid; Blue light...Addgene's guide to using Optogenetics plasmids in your lab, including explanations of microbial opsin...Addgene, check out our Optogenetics Plasmid Collection . Microbial Opsins Opsins are light-gated ion channels... of opsins here to find the right optogenetics plasmids for your experiments. Channelrhodopsins Channelrhodopsins...SwiChRca, Phobos, Aurora Browse Channelrhodospin plasmids . Halorhodopsins Halorhodopsins are light-gated...
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