... that (1) the sequence is unique compared to the rest of the genome and (2) the sequence is located just...

.... Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent cells do not...establish cell lines that stably express a gene of interest. While the general approach for using lentivirus...the positively charged polycation, polybrene. Interestingly, one study found that alternative polycations...

...well as plasmids that already contain a region of interest before or after luciferase . In addition, Addgene... pGL3-LRR : Genomic tiles spanning a region of interest are cloned into a Firefly luciferase reporter ...you can clone your regulatory element or gene of interest into to create a luciferase reporter. Unless otherwise...Lentiviral expression of firefly luciferase and gene of interest Bryan Welm 18760 MSCV IRES Luciferase Firefly ...Retroviral expression of firefly luciferase and gene of interest Scott Lowe 18782 MSCV Luciferase PGK-hygro Firefly...expression of firefly luciferase and dTomato. A gene of interest can also be inserted into the plasmid. Malcolm...construct that contains your regulatory element of interest. ID Plasmid Description Gene/Insert PI Return ...

...Enzyme Collection DNA Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases...polymerases, DNA ligases, reverse transcriptases, and restriction enzymes. These enzymes are useful in both basic...the links below: DNA Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases...-phi29pol phi29 DNA Polymerase Return to top Restriction Endonucleases Addgene ID Plasmid Gene/insert ...

...acids (ncAAs). A plasmid containing the gene of interest with the modified codon (typically the amber codon...a non-canonical amino acid into the protein of interest: The non-canonical amino acid, which is generally...codons, this can be challenging. In E.coli , the rarest codon is the amber stop codon (UAG) and thus this...this codon is often used. The gene of interest can be expressed from a plasmid containing a UAG codon ...sites changes to UAG, RF1 function removed, MutS restored so decreased mutation rate George Church 68306...

... incorporation phosphoserine into your gene of interest....choosing plasmids to express the phosphoprotein of interest, a medium copy RSF1030 origins or a low copy p15a...domain and the phosphosite, mCherry fluorescence is restored, so interactions can be discovered by fluorescence-activated...identify candidate human substrates of kinases of interest by mass spectrometry in a technique called serine-oriented...

... The following resources may be of interest for groups interesting in implementing intersectional experimental...ssitrategies to define cellular sub-populations of interest, including dual-transgenic recombinase-expressing...testing of novel INTSRECT plasmids for groups interested in producing their own INTRSECT plasmids. Online...

...step 3.1.4 ), 5.0 μl restriction enzyme buffer (10x), 4.0 μl (20 U) BbsI restriction enzyme (5,000 U/ml)...for cloning into the pX330 vector using BbsI restriction enzyme ( Table 3 ). Add a G nucleotide after ... construct. Transfecting CRISPRs into Cells of Interest NOTE: This protocol involves the delivery of CRISPR...

...Plasmids for REAL (Restriction Enzyme And Ligation) TALEN assembly containing single TALE repeat domains...any desired sequence, referred to as the REAL (Restriction Enzyme And Ligation) assembly method ( Sander...encoding a single TALE repeat domain and flanked by restriction sites used for ligation into longer arrays. These...

...between cancer cells, therapeutic drugs, and the rest of the body than studies in other model organisms...level and spatiotemporal expression of a gene of interest. This type of control over gene expression is ...continuously improved upon since then. If you are interested in these systems check out Gerald Rubin’s lab...specific plasmids (L4440) that contain your gene of interest (Fire et al., 1998). The discovery of RNAi in ...sensor, GCaMP6 (Nguyen et al., 2015). If you are interested in C. elegans plasmids and resources check out...

...release the plunger, allowing it to go back to its resting position. Wait for the liquid to stop flowing into...

...it's important to determine where your protein of interest resides. There are a variety of methods for determining...to compare the localization of your protein of interest with known markers. Addgene has assembled a collection...subcellular structures. You should fuse your protein of interest with a fluorescent protein that has a different...localization or possible function of a protein of interest. Browse or search the table below to find markers...

...best-characterized effects of p53 are in promoting cell cycle arrest, apoptosis, or senescence in damaged cells. The... receptor Gadd45 GADD45A GADD45B GADD45G Growth arrest and DNA-damage-inducible; alpha, beta, or gamma...BCL2 binding component 3 Reprimo TP53 dependent G2 arrest mediator candidate Scotin Shisa family member 5...

...promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also ...transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding...specific promoter or enhancer for your gene of interest. Available modifications include histone acetylation...

...encode a wide range of gene products. The main restrictions on a lentiviral transfer plasmid are size (i.e... Any expression cassette (promoter and gene of interest) can be cloned into the plasmid. Nolan 12257 pWPXL...bi-cistronic expression of EGFP and the gene of interest, also can express shRNA (cloning H1-shRNA cassettes...

...plasmids for endogenous tagging of your gene of interest.... design. How to use CRISPR to tag your gene of interest Mendenhall and Myers Tagging System The Eric Mendenhall...plasmids as listed in each row. If your own gene of interest is currently unavailable, you will need to design...

...out my window and I saw the Dorr Mountain with the rest of the skyline of Acadia National Park. I was confused...molecular biologist with Biogen and is particularly interested in applying CRISPR for mouse genome editing. ...Zoology, Chinese Academy of Sciences, and he is interested in using CRISPR to develop therapeutic applications...

... the experiment, preventing contamination of the rest. Avoid long exposure of the cell culture to air ...

... monoclonal cell lines by limiting dilution. The rest of the cells were frozen down.     Pro-Tips: 1. ...

...control of nanos promoter and 3'UTR. For germ line restricted genome engineering in Drosophila melanogaster...control of nanos promoter and 3'UTR. For germ line restricted high specificity genome engineering in Drosophila...