We narrowed to 432 results for: sequence

... and then plasmid sequencing if you can. To quote Doench, “Whole plasmid sequencing is just part of doing...enzyme that cleaves outside of its recognition sequence. This means that each component can be designed...? The Doench lab tested the Fragmid system by sequencing 60 individual assemblies created over five months...82 of the correctly banded clones for plasmid sequencing and found 98% of the plasmids were perfectly ...confirm that at least one plasmid was the correct sequence for every individual assembly. What destination...051 (for stable fly cell lines; attL and attR sequences for genome integration by phiC31) Drosophilia...pRDB_053 (for transgenic flies; gypsy insulator sequences; white + selectable marker; attB site for genome...

...next-generation sequencing (E). Considerations and tips for successful screens Next-generation sequencing - CRISPR...guide RNA (gRNA). The ~20 nucleotide targeting sequence within the gRNA is defined by the user, and it...immediately 5’ to a protospacer adjacent motif (PAM) sequence. Co-delivery of wild-type Cas9 and a gRNA generates...screening due to the broad range of potential target sequences and ease of generating gRNA-containing plasmids...generate lentivirus. Be sure to use next-generation sequencing to check the quality of your amplification. If...by a unique barcode on each plasmid. As such, sequencing CRISPR libraries after amplification and after... a screen requires the use of next-generation sequencing. Representation - Most libraries contain 3-6 ...

...machines (RSMs), which take on a particular DNA-sequence configuration (state) based on the identity and...RSMs work? The state of an RSM is defined by the sequence within a prescribed region of DNA termed the “...straightforward way to read the state of an RSM is to sequence the register. In our paper (1), we allowed individual... register region and sent the PCR product for sequencing.  A less straightforward, but more population-wide...of state interrogation mentioned above (Sanger sequencing, qPCR, and fluorescent reporter-based) each come...with their advantages and disadvantages. Sanger sequencing is simple and the most accurate method as it ...the state of a register. However, each Sanger sequencing reaction can only be performed on one register...

...transcription of RNA such as long-read cDNA sequencing and RNA sequencing of complex heterogeneous RNA samples...functional element (tyrosine sulfation consensus sequence, fluorophore, or APEX2), allowing for quantitative...Caspex’ construct is guided by a pre-defined sgRNA sequence to a specific genomic locus and labels the proteins...radius of approximately 20 nm of a specific DNA sequence in only 1 min. Using these lentiviral plasmids...Shield1 inducible CMV promoter and targeted to a sequence of interest with an sgRNA expressed from a separate...

...that remained unexplored. Using new single-cell sequencing approaches, the lab was able to eliminate the...cultures to study these microbes. From 16S rRNA sequencing, hot springs were known to be diversity hotspots... characterized a few samples by 16S rRNA gene sequencing which showed the presence of many “dark matter...samples, and years later, several whole genome sequencing projects were launched in the Quake Lab. Identifying...mini-metagenomics method using microfluidic parallelization to sequence this highly complex hot spring sample. The sample...separated into smaller samples of 5-10 cells each for sequencing, allowing for lower complexity with higher throughput...

...and amplification of a recombinant DNA without sequence alteration. Molecular cloning techniques are some... DNA polymerase, the construction of specific sequences for base pair overhangs, insert and vector purification...products, nor does it require the use of specific sequences. Read on to learn how easy this process is and.... The primers used to amplify the insert have sequences specific to the insert with an additional 15-17...with the appropriately sized product, purify and sequence the plasmid to confirm no mutations were introduced...FastCloning: a highly simplified, purification-free, sequence-and ligation-independent PCR cloning method." ...

...Deleting a large region of the genome by removing the sequence between two target sites. Modifying multiple genes... oligonucleotides for your chosen gRNA target sequences and clone them in just as you would for a single...enzymes, which cleave outside of their recognition sequence, creating flanking overhangs. These overhangs ...the oligonucleotides specifying each gRNA target sequence into distinct expression vectors using the enzyme...the promoter-gRNA construct, but with different sequences adjacent to the sites. When digested with the ...Type IIS enzyme, the unique flanking overhang sequences can link together to allow for ordered assembly...schematic below.   Figure 2: gRNA target sequences (colored rectangles) are cloned into various plasmids...

... the Cas12a-sgRNA complex binds to the target sequences, Cas12a activity gets activated. This unleashes... of interest. Cas13 uses a complementary crRNA sequence to bind to the sample and upon binding, gets activated...trans cleavage reaction that cleaves a nearby RNA sequence that contains a fluorophore that is quenched when... Unlike Cas12a, Cas13a does not require strict sequence preferences at the target site, while Cas12a requires...SHERLOCK diagnostic test, this test varies in the sequence of steps and the Cas protein used. Here, all the...to one strand of the reverse-transcribed target sequence that are present close to the primer recognition...binds to the complementary region in the target sequence and this activity results in the trans cleavage...

...primers that contain the appropriate homologous sequences. NEB recommends an overlap of 15-40 bp, with a... insert both the gene of interest and the tag sequences into the vector in one step without scars as depicted...might be anticipated to occur within a terminator sequence), since this would directly compete with the required...assembling multiple fragments in one step with no scar sequences remaining in the final product. The long overlapping...fragments as compared to the smaller overlapping sequences left with a restriction digest. A study in 2013...around this is to use a combination of standard sequences with overlap regions, such as in the case of MODAL...overlap-directed methods (Casini 2014). In this case linker sequences are added before the homology regions, which allows...

...questions, biologists often use next-generation sequencing. Sequencing the 16S rRNA reveals the identity of the...the organisms present while sequencing of all transcripts gives clues into what the microbes are doing...doing. But aside from sequencing, scientists can also study the microbiome by using engineered genetic tools...While the ability to monitor the microbiome with sequencing is vast, the ability to genetically alter microbiome...biomarkers In clinical and diagnostic labs, deep sequencing and qPCR methods are too costly and time intensive... be designed to bind and detect nearly any RNA sequence. Downstream of the hairpin is a ribosome binding...