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

...where you are going and what you expect. For the rest of the chalk talk, as with your job talk, avoid ...and tissues is fundamental to development. I'm interested in how cells find their way to the correct positions...expert in [your research area]," – you should show interest in other faculty members’ research. Still a Grad...Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory proteins...

...conditions:  1) The sequence is unique compared to the rest of the genome.  2) The target is present immediately...Cas9-mediated frameshift mutations in the gene of interest. The genomic target for Cas9 and the introduction...site early in the coding region of the gene of interest. This gives you loads of sequences to select from...to find a SpCas9 compatible PAM at your site of interest, that is the most efficient option. If there is...repressors to the promoter region of the gene of interest. dCas9-based repressors are useful when knocking...

.... 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...

...Express gene of interest from the CMV promoter pCDH-EF1 72266 Express gene of interest from the EF-1 promoter... Express gene of interest from the CB promoter pCDH-PGK 72268 xpress gene of interest from the PGK (phosphoglycerate...Express gene of interest from the CB promoter pCDH-EF1s 72484 Express gene of interest from a truncated...respectively, and empty vectors to express your gene of interest under the control of a variety of promoters. Professor...kinase I) promoter pCDH-CMV4 72284 Express gene of interest from the CMV promoter. WPRE has been removed to...

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

...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 ...

...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 ...pEVOL-NBK-1 PylRS M. mazei Pyrrolysine Bacterial Cole DeForest 209738 pAB228i16 G1 PylRS Methanogenic archaeon...sites changes to UAG, RF1 function removed, MutS restored so decreased mutation rate George Church 68306...

... easily gate on the unlabeled cells of interest while the rest can be excluded, or “dumped”. From left...fluorophore-conjugated antibodies directly bind to epitopes of interest that are found in the extracellular space. The...like bulk lympho-/splenocytes, you might only be interested in one of the various cell populations; for example...selection: labeling markers that your cells of interest do not have, and selecting for the cells that ...quick method of negatively selecting your cells of interest is magnetic bead-activated cell sorting, or MACS...An effective strategy to sort out your cells of interest via FACS using negative selection is called dump...Intracellular labeling Oftentimes, a marker of interest is not located on the cell membrane but inside...

...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...

...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 ...

... 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...strategies 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...

...various fluorescent markers and tag your proteins of interest with fluorescent proteins in yeast.... overexpression of FPs and tagging of genes of interest with FPs in yeast. Characterization of some of...tagging vectors to create fusions of proteins of interest with yeast codon-optimized fluorescent proteins...

...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...