Skip to main content

We narrowed to 605 results for: RAN-1

Showing: 261 - 270 of 605 results
  1. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...homologies. Abbreviated protocol for PITCh Step 1: Generate microhomologies in the PITCh vector ~20...editing possibilities MMEJ can enable. References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted ...CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids from ...genome edits. But have you heard of another double-stranded break repair mechanism: MMEJ (microhomology-mediated...primary methods for repairing DNA after a double-stranded break. HDR copies the sequence from a repair template...are chewed back to reveal homology, allowing the strands to anneal. DNA synthesis then fills in the gaps...5-25 bp) regions of homology flanking a double-stranded break yield precise deletion of the sequence between...
  2. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...biological questions. The two most common uses are: 1) to test the expression level in a specific system...brightness is compared to that of EGFP which is set as 1. Some proteins are very dim (e.g. TagRFP657, which...Photostability can be as short as 100ms (EBFP) or as long as 1 hour (mAmetrine1.2). However, for most FPs it is a...or Förster] resonance energy transfer). FRET occurs when energy transfer from one FP (e.g. CFP) excites...classified by the emission color (emission wavelength range) FPs are usually classified by emission color as...as outlined below (or emission wavelenght range).  By mutating GFP, the variants blue FP (BFP), cyan FP...nm Green 499 - 519 nm Yellow 524 - 538 nm Orange 559 - 572 nm Red 574 - 610 nm Far-Red 625 -...
  3. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ... spread out onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body ...a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...growth rate/transfection efficiency is not known, this is easily achieved by splitting transfected cells into...specific cell line, and its transfection efficiency. We often trypsinize the transfected cells on the day after...after transfection into plates containing selection agent at different densities, then dispose of the plates...under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to the gene of interest ...such as the Viaflo from Integra Biosciences) to transition from a 48-well cell plate to a 96-well assay ...
  4. Editor's Choice, September 2016

    Type
    Blog Post
    Published
    Oct. 7, 2016, 4 p.m.
    ... products). I picked this post for a few reasons: 1) I think phage are super cool and have considered ...Connect the Research Community Grad School Advice Part 1: Picking a Lab and a Project That’s it for now. As...Editor's Choice Blog Posts As I’m sitting in the San Francisco International Airport listening to the Lion King...back from a very successful Outreach Trip in San Francisco and would like to give my thanks to all the Addgene...
  5. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology.     Fig. 1: SpCas9 has sparse PAM coverage in the human genome.... The activity of SpRY was validated in double stranded cleavage and base editing applications, with high...solution? Almost! Assuming a PAM of NRN, 50% of randomly selected genomic loci will have a PAM located ...
  6. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    Published
    July 7, 2017, 1:32 p.m.
    ...Listen to the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University...Unit, the Pulmonary Consultation Service, the Alpha-1 Center, and the Pulmonary outpatient clinic, all at...Chapter 3: How all basic research ends up being translational. Ex: the discovery of GFP. Chapter 4: Goals ...
  7. Plasmids 101: Colony PCR

    Type
    Blog Post
    Published
    May 12, 2016, 2:30 p.m.
    ... for less money. The key steps to colony PCR are: 1) design primers to detect the presence of your insert...primers. There are 3 strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers...previously described. When using insert-specific primers (1), positive clones (+) will give a band, while a negative...control. A good positive control is bacteria transformed with the same backbone plasmid. If this control...strain. A good negative control strain is an untransformed culture of the same strain of bacteria you used...Cloning Technique Learn to Perform a Bacterial Transformation Deposit Your Positive Clones with Us ...
  8. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    Published
    July 23, 2019, 12:46 p.m.
    ...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6...PHP.B has a >40-fold increase in CNS transduction and transduces the majority of astrocytes and neurons... mice whose CNS was transduced by PHP.B but absent in mice who were not transduced by PHP.B. The Deverman...These results were surprising because in vivo transduction of AAV serotypes generally tracks between strains...diverse library of AAV9 mutants that had a small randomized peptide loop inserted in the AAV9 capsid. This... an enhanced version of PHP.B, shows similar transduction efficiency, but at a log lower dose than PHP.B...
  9. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...individual activities (1). Researchers also discovered the modular format of some transcriptional activators: that...requirement for transmembrane proteins, a split ubiquitin system has been devised (8). References 1. Casadaban...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...flurry of discovery in the field of eukaryotic transcriptional activation and cell biology. During this period... that the DNA binding domain (DBD) and transcriptional activation (TA) domains retain their individual... regardless of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists...interactions. In later years, improvements to yeast transformation protocols also helped establish the yeast two-hybrid...
Showing: 261 - 270 of 605 results