... mutagenesis vastly facilitates the subsequent process of screening for succesfully mutated clones. Forward... introduce undesired mutations through the PCR process, you need a high-fidelity polymerase. There are...fragment length polymorphism (RFLP) analysis. In this process, if your mutation introduces an additional restriction...DNA duplex stability from the base sequence." Proceedings of the National Academy of Sciences 83.11 (1986...Yongzhen, et al. "New insights into the QuikChangeTM process guide the use of Phusion DNA polymerase for site-directed...

...doing an experiment wrong. However, even simple procedures like pouring a plate or running a gel have intricacies... retain searchable, thorough notes on how lab procedures were actually performed, potentially curtailing... sense on many different levels - it’s hard to process large amounts of information and, from a SciComm...how and why papers change throughout the review process. This will hopefully push reviewers towards more...experimental design as part of the publication process (Nosek et al 2018). In this ideal situation, a ...

...generation easi-er! Generating mice before CRISPR The process of generating knock-in mice pre-CRISPR was very...would be unsuccessful. CRISPR made the overall process of mouse model generation simpler, as described...knockout alleles or point mutations, the 8-10 month process of conventional gene targeting is lowered to about...embryo to create chimeric mice. This laborious process takes ~8-10 months. With CRISPR (B), you can inject...than i-GONAD and is a more technically difficult process. Overall, the main drawback of i-GONAD is that ...

...this blog post, we'll discuss the principles and process of designing a flow panel.  Principles of excitation...electrons within atoms to a higher energy state. This process is called excitation. After a short period, the...spectrum compared to APC. This happens due to a process called Fluorescence Resonance Energy Transfer (...constantly happening in cell culture, the transduction process itself also leads to some cell death. As a result...be absorbed and emitted by matter, and how this process creates visible colors. Next, you learned about...

...specific subset of cells in a population in a procedure termed fluorescence activated cell sorting or ...forces the cells to line up in single file in a process termed hydrodynamic focusing. This allows each ...factors including sample preparation, staining procedures, and controls. To prevent instrument clogs, ensure...control with cells that went through the staining procedure minus the antibody. An unstained control allows... to aid with compensation. Compensation is the process of correcting spectral overlap that occurs between...

...the Red2Green processing facility, where it’s sanitized through an EPA-approved process and no longer ...focused on streaming some lab waste for recycling- a process that’s integrated into waste management in our ...we consider the environmental impact of those processes? Who even regulates them? You may be surprised...

...capability with the endogenous tRNA-processing system.” Proceedings of the National Academy of Sciences...short answer is no. Unless you use a system for processing a continuous multi-gRNA transcript, each gRNA... Note: depending on which method you use, the procedure will vary slightly.   Two Golden Gate options...and somatic genome engineering in Drosophila.” Proceedings of the National Academy of Sciences USA 111(29...

...Direct reprogramming describes the process where differentiated cells are turned into a cell type of ...medicine, direct reprogramming is an inefficient process, with the majority of cells failing to develop ...expression of transcription factors Foxa1 and Hnf4α, a process that like other reprogramming protocols is inefficient...CellTag could be used to study other biological processes where cells are undergoing identity changes, such...

... Hot Plasmids Podcast!  MarathonRT: A highly processive reverse transcriptase Article contributed by ...several important shortcomings including low processivity (ie. the probability of the polymerase continuing...heterogeneous RNA samples. Furthermore, such high processivity enzymes and the structural understanding thereof...to existing technologies based on polymerase processivity. Please find the MarathonRT plasmid here! Zhao...

... planning, visualizing, and documenting their procedures. Preventable errors in the design of cloning ...of software with these qualities is an ongoing process that involves iterative refinements in response... algorithm and thereby speed up the annotation process. Many of the best ideas come from engaging with...use SnapGene during the plasmid quality control process Check out full plasmid sequencing at Addgene Additional...

...sample and target type. To start the validation process, one must first confirm that a candidate antibody...approved antibody clones The antibody validation process represents a high level of effort and time, which.... We find it useful to start with a screening procedure in easily genetically modified cell types (top... C. (2023). Scaling of an antibody validation procedure enables quantification of antibody performance...Implementation of an antibody characterization procedure and application to the major ALS/FTD disease gene...

...systems have a variety of enzymes responsible for processing foreign DNA as well as the RNA guides required...scaffold linking the crRNA to Cas9 and facilitates processing of mature crRNAs from pre-crRNAs derived from...repeat regions contain sequences required for processing pre-crRNA into mature crRNA and tracrRNA binding... Overview of native CRISPR arrays and their processing for cleaving foreign DNA. A) CRISPR arrays as...

...came to MIT to interview, part of the interview process was several short talks by current faculty members... it was just this complete black box. The RNAi process had just been discovered, the mechanism was barely...what confers specificity in the miRNA regulatory process. This early research is still relevant to his work...determining what genes are involved in biological process X, you can do with CRISPR technology...” and “with...

...is unlikely to be caught during the peer review process, which often focuses more attention on the results...noise There is, of course, simple noise in the process of sampling and inference-making. Even perfect ...exist or are preserved as part of the publishing process and become lost if not archived. Remove bias with...with preregistration Preregistration is the process of creating a time-stamped research plan in a public...

...mutation allowing for a functional enzyme. This process is called α-complementation. The system described...are working as they should be. Don't rush the process: It is important to give your plates enough time...any intact β-galactosidase to be expressed and process x-gal into blue pigment (16-20 hours). A plate ...Blue-white screening is just that – a screening process. It does not select only those cells that have ...

... as part of their infection and/or replication process within the host. dsRNA viruses are inherently double...intermediates as part of the viral replication process. dsDNA viruses yield dsRNA through a slightly different...inherently generating RNA:DNA intermediates in the process. These transcription-coupled R-loops are transient...above play distinct roles in promoting cellular processes and functions, but they also have a darker side...

...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9...stranded DNA degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction...) and does not require a tracrRNA (it can self-process pre-crRNA into crRNA). The size and simplicity ...11 cuts ssRNA and is capable of pre-crRNA self-processing. Functionally, it is very similar to Cas13 and...

...helps me see problem solving as a generalizable process.  Step One: Identify the problem and gather ...fix everything or revealed another issue in the process, you might need to go back to the beginning, reassess...strategy. Step Five: Share your results.  Does this process remind you of anything? Perhaps the Scientific ...be able to carry out the entire problem solving process on your own, but you can start by paying attention...

... at the mutation site, indicating that repair proceeded through NHEJ rather than HDR. Figure 1:...: Homology-directed repair in targeted zygotes proceeds from the maternal allele rather than the supplied... characterization studies, the edited embryos proceeded to the blastocyst stage at similar rates to untreated...surrounding such research, it is crucial that scientists proceed with caution and with proper oversight from regulatory...

...want to mentor you during the grant application process. When my PhD advisor first asked me to write a ...PI asks for assistance in the grant application process, she has faith in you! Additionally, keep in mind...motivating research will help you understand the process and continue to improve your grant writing skills...Still want more information on the application process? What types of writing feedback do you find most...