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We narrowed to 82 results for: total dna

Showing: 11 - 20 of 82 results
  1. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    ...transformed with the library DNA, they grow and replicate, and then their DNA is harvested, much like one...However, lots of library DNA is needed for screening experiments, so the scale of the DNA prep is larger. And...eight electroporation reactions using a total of ~400ng of library DNA. These reactions should allow us to...pelleted for DNA extraction. Prepare library product for next-generation sequencing Once the DNA has been ... library during transformation First, use enough DNA and competent bacteria to ensure all components of..., skewing the composition of the final recovered DNA. This bias can be counteracted by growing the cells...NGS samples using both pre- and post-amplification DNA and generates PCR sequencing products from both. ...
  2. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    ...identified a set of one to three mutant complementary DNAs (cDNAs) representing core nodes that, when over-expressed...cloned all the cDNAs into a single consensus lentiviral expression vector and attached a DNA barcode to each... things - sometimes drug screens and sometimes totally different experiments we never thought of - and...curate the perfect library, obtained the appropriate cDNAs from other researchers, and cloned them into lentiviral...
  3. Learning Technical Skills at a Lab Central Workshop

    Type
    Blog Post
    ...NEBNext Ultra FS DNA Library Prep Kit To create a DNA library, one must first fragment a DNA sample into many...about high throughput DNA prepping by trying out NEB’s new NEBNext Ultra II FS DNA Library Prep Kit for...Addgene, part of my job, in addition to preparing DNA and generating/replenishing kits, is to find new ...working on a project where we aim to do large scale DNA preps with multiple samples without cross contamination...components. Without proper processing, these small DNA strands can form concatemers in later steps. This... NEB kit includes a step that both fragments the DNA and prevents it from forming concatemers later. The...released prep kit. In their prior NEBNext Ultra II DNA Library Prep Kit for Illumina, one was only able ...
  4. Let There Be LITE Plasmids

    Type
    Blog Post
    ...Feng Zhang’s lab at MIT, the use of customizable DNA-binding domains based on transcription-activator-...established for conventional optogenetic tools." In total, the researchers applied the LITE system to 28 different...
  5. RNA Extraction Without A Kit

    Type
    Blog Post
    ... the intact RNA from DNA and proteins. Although very effective at isolating total RNA, this method requires...precipitate RNA, but not DNA and proteins. Lastly, using a DNase (there are several DNase enzyme products on...As with DNA isolation, scientists commonly rely on RNA isolation kits to make their life easier. Recently...Recently, we published a blog on DNA purification without a kit that outlined several reasons why doing ... one should take care whenever doing any type of DNA or RNA purification to avoid contamination, take ...RNA extraction. RNA is inherently not as stable as DNA- it is single stranded and its ribose groups are ...There are many alternative methods for isolating DNA without a kit. However, that isn’t the case for RNA...
  6. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...thermophilus to predatory phages to test if the exogenous DNA would be incorporated into the bacterial genome. ...specifically, specialized Cas proteins snip the foreign DNA into small fragments approximately 30bp in length...Once the CRISPR/Cas complex binds to the foreign DNA, a cut is made to destroy the invader. Wikipedia...slightly different Cas proteins and PAM sequences. A total of eight evolutionarily conserved CRISPR subtypes...
  7. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...sub-culture, and prep with a DNA prep kit or your lab's favorite in-house protocol. DNA yields for this procedure... levels of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein expression... improved plasmid DNA yield. Culture conditions play a critical role in plasmid DNA yield. Different plasmids...typically in excess of 100 ng/ul, more than enough DNA to verify your plasmid via sequencing or restriction...restriction digest. Chances are, you’ll even have DNA left over for other applications, like PCR, cloning, transfection...use NEB Stable to transform and prep high quality DNA that contains these elements. Be aware of endA+ strains...endonuclease that can sometimes co-purify with your plasmid DNA and lead to degradation of your prep. Many plasmid...
  8. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...20 ng of total gDNA is sufficient for analysis. We have routinely successfully analyzed gDNA from a few...will contain either one or no copies of the target DNA. Conventional thermal cycling is then performed to...Figure 1). Partitions that did not contain any target DNA at the onset of PCR are negative, whereas those that...be used to determine the actual number of target DNA molecules present in the original sample with a great...mutation in a bulk or single cell-derived genomic DNA sample as part of a precision genome editing workflow...a great screening tool, it still requires target DNA sequencing to confirm the exact nature of your mutations...
  9. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged DNA is extracted from...into longer DNA contigs without using a reference sequence. The five contigs with the most total reads are...used to identify DNA contaminants in AAV preparations and provide information on these DNA species with exquisite...increases. In addition to human error, fragments of DNA from the helper plasmids or the cell genome can be...determine the identity and serotype of the packaged DNA and look for potential contaminants. The analysis is ...reviewed. As mentioned earlier, it is common to find DNA from the packaging cell genome, bacterial genomes...68544 to validate our method. We then extracted the DNA and processed the mixed sample in our pipeline (see...
  10. Lab Automation at Addgene

    Type
    Blog Post
    ...rate! Automating DNA purification We're developing ways to prep large quantities of DNA because some countries...more efficient. Here, I’ll discuss transformation, DNA purification, and filling vials with agar. I’ll also...Biomek. The FXP id also able to do high throughput DNA preps and other molecular work if we need it to. ...globe, we’re working on ways to send plasmids as DNA as opposed to in live bacteria. For these shipments...shipments, we’ve needed to find a way to purify DNA from many bacterial samples at the same time as opposed...aliquoting. Simply program the pipette with how much total sample you need and the volume of the smaller aliquots...
Showing: 11 - 20 of 82 results