...OZ551 and OZ552 OPEN OPEN gGTCATCTGCcattcGAGGCGGACa REST corepressor 2 OZ553 and OZ554 OPEN OPEN gCTCACCATCtgttggaTGCGATGGAa...

...Adult Dermal Fibroblasts Neurons Lentiviral Human REST suppression mediates neural conversion of adult ...

...conditions: The sequence is unique compared to the rest of the genome. The target is present immediately...sequence is for the genomic target compared to the rest of the genome. Ideally, a gRNA targeting sequence...nucleotide edits that introduce a novel restriction site. The region of interest is PCR amplified, digested with...region of interest can be PCR amplified using primers that (A) flank the region of interest (deletions...sequence must be delivered into the cell type of interest with the gRNA(s) and Cas9 or Cas9 nickase. The...identify are actually important for your phenotype of interest. You can individually test the gRNAs identified...to ensure that they reproduce the phenotype of interest. Find more information on factors to consider ...

...Stathmin-2 Rat Mouse IgG2a 140066 Anti-Co-Rest/RCOR1 [K72/8R] Co-Rest/RCOR1 Human Mouse IgG2a 140068 Anti-Kv1.6...

...protein of interest in E. coli , B. subtilis and P. pastoris . All plasmids use the same restriction enzymes...relatively well understood biology to answer many interesting questions. Does my transcription factor bind ...and/or alter cellular activity in a way that’s interesting from an applied perspective, sometimes it’s best...Plasmids Tn7 Transposition Nancy Craig A gene of interest cloned into one of these plasmids can be transposed... many enterobacterial genomes. The bacteria of interest are transformed with the appropriate construct...get a better understanding of what your gene of interest does when expressed in vivo . Plasmid ID Visualization...fluorescent protein fusions with your protein of interest. See an overview of Addgene fluorescent protein...

...protein of interest. Signal strength is determined by how many antibodies bind the protein of interest, while... antibody of interest to a second, independent antibody for the same protein of interest, with no overlapping...quantify, identify, separate, and locate proteins of interest. The protein that an antibody recognizes is called... of the animal’s immune system. The antigen of interest is introduced to the animal, which then mounts...primary antibody which is bound to your protein of interest. Secondary antibodies can be isotype-specific ...recognize an available epitope on your protein of interest. Polyclonal antibodies are typically collected...Western blots should be run with your protein of interest and at least one ubiquitously expressed protein...

... of interest fused to the N-terminus of Amber Amber C1 Yellow Mammalian Express a gene of interest fused...a fluorescent fusion protein with your gene of interest and the listed fluorescent protein. Plasmid Color... mCerulean N1 Cyan Mammalian Express a gene of interest fused to the N-terminus of monomeric Cerulean ... mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric Cerulean ...His tag SCFP3A Cyan Mammalian Express a gene of interest fused to the C-terminus of SCFP3A Amber N1 Yellow...Amber mVenus N1 Yellow Mammalian Express a gene of interest fused to the N-terminus of monomeric Venus mVenus...mVenus C1 Yellow Mammalian Express a gene of interest fused to the C-terminus of monomeric Venus pCEP4YPet-MAMM...

...-CDS issues. Prohibited Restriction Sites: EcoRI, XbaI, SpeI, PstI Restriction Sites to Avoid: NotI For...Coli genome. Prohibited Restriction Sites: EcoRI, SpeI, NheI, PstI Restriction Sites to Avoid: PvuII, ... use of assembly standards – combinations of restriction sites which preserve a standard prefix and suffix...backbone can be repeated to ligate your gene of interest after the promoter, and so on. The following table... reverse complimented for primer design. The restriction enzymes used for assembly are the prohibited ... Employs high-efficiency enzymes. Prohibited Restriction Sites: EcoRI, BglII, BamHI, XhoI For more info...giving in-frame scar encoding Thr-Ala. Prohibited Restriction Sites: EcoRI, XbaI, SpeI, PstI For more info,...

...Experimental Tips Choosing a tet system If your gene of interest should be active, and only turned off occasionally...off or tTA is more appropriate. If your gene of interest should be mostly inactive, and only turned on ...tetracycline-dependent promoter upstream of your gene of interest tTA expression plasmid Components for a tetracycline...tetracycline-dependent promoter upstream of your gene of interest rtTA or TetR expression plasmid Stable cell lines...will allow you to insert and induce your gene of interest. ID Plasmid Description Co-expressed tTA, rtTA... 16542 pBI-MCS-EGFP Expression of your gene of interest (MCS with a β-globin poly A) & EGFP from a bidirectional...plasmid for inducible expression of transgene of interest and EGFP None Either Lung 11662 pPRIME-TET-GFP-FF3...

...analysis to purify specific genomic regions of interest 48807 8xLexA-binding elements/pMD20 iChIP plasmid...analysis to purify specific genomic regions of interest 51258 3xFLAG-dCas9/pMXs-IG Expresses 3xFLAG-dCas9...analysis to purify specific genomic regions of interest 51259 3xFLAG-dCas9/pMXs-I2 Expresses 3xFLAG-dCas9...analysis to purify specific genomic regions of interest 51260 3xFLAG-dCas9/pMXs-neo Expresses 3xFLAG-dCas9...analysis to purify specific genomic regions of interest 61079 gRNA-hIRF-1 #12 Expresses a guide RNA (gRNA...analysis to purify specific genomic regions of interest 63589 3xFN-Tel-TAL/pCMV-7.1 Expresses a 3xFLAG-tagged...analysis to purify specific genomic regions of interest 82613 3xFLAG-dCas9/MSCV-EGFP Expresses 3xFLAG-...

...can precisely control expression of your gene of interest. Many Cre constructs also contain fluorescent ...a promoterless vector with several available restriction sites none Mammalian Sauer 11957 pBS595 tet-hCMV-EGFPcre...with stop codon Entry vector Ko 139546 pENTR-CreStopDeleted Cre without stop codon Entry vector Ko 139547...-lox” or “LSL” cassette) upstream of a gene of interest will prevent gene expression in the absence of... Switch: These constructs contain two genes of interest, Genes A and B. When Cre is absent, only Gene ...and type of target sites that flox your genes of interest makes FLEx switch a powerful experimental tool...information. Empty backbones to regulate your gene of interest: ID Plasmid Description Expression System PI 11578...

...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...

...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...

...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...