...plants. Further opportunities with pSiM24 While these properties of pSiM24 are suitable for many applications... of DNA that will be transferred to the plant, similar to viral vectors’ terminal repeats. It’s a lot ...larger cloning capacity would be very useful: enter pSIM24! To build a better binary vector, Sahoo et al. ... Sahoo et al. demonstrate that this new vector, pSiM24, can also be used with biolistic methods, making...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available...blotting. Given its versatility and customizability, pSiM24 can be used not only for plant engineering and ... Kumar, Nrisingha Dey, and Indu Bhushan Maiti. "pSiM24 is a novel versatile gene expression vector for...

...can be used in combination to simultaneously perform modifications, simply by adding all the necessary ...allow any cloning procedure to be performed using a simple two-step, 2 hr protocol prior to transformation...linear fragments need to get into the same cell. For simple cloning requiring just one or two recombination...Subcloning - To move a gene from one vector to another simply amplify the vector and insert with separate primer...colonies. We routinely screen just two colonies for simple cloning procedures. Use high fidelity DNA polymerase...

... cloning techniques with higher efficiency and simpler procedures have been developed. Some of these include... excessive time requirements. FastCloning is a simpler yet reliable cloning technique that was developed...2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and ligation-independent...

...treatment can photoconvert GFP from green to RED! This simple technique has been shown to work in plant, Drosophila...

...can bring down the morale of a group with their pessimism, Know-it all experts, who think their way is...

... have no fear - I’m here to introduce you to a simpler, yet more effective, CRISPR-based epigenetic approach...Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving...blasticidin or puromycin resistance genes.  It’s that simple! Good luck and happy editing!    Thank you to...

...Mammalian TAG Simon Elsaesser 140011 pAS_4xMx PylT_FLAG-Mx PylRS PylRS M. alvus CpK Mammalian TAG Simon Elsaesser... TAG Simon Elsaesser 140013 pAS_4xMx PylT C41CA_FLAG_Mx PylRS PylRS M. alvus Mammalian TAG Simon Elsaesser...TAG Simon Elsaesser 140023 pAS_4xMma PylT_FLAG-Mma PylRS AF PylRS AF M. mazei Mammalian TAG Simon Elsaesser...Mammalian TAG Simon Elsaesser 154763 pAS_FLAG-G1 PylRS G1 PylRS M. archaeon Mammalian TAG Simon Elsaesser ... TAG Simon Elsaesser 154768 pAS_4xG1 PylT FLAG-G1 PylRS G1 PylRS M. archaeon Mammalian TAG Simon Elsaesser...Mammalian TAG Simon Elsaesser 174896 pAS_4xMmaPylT(UUA)_EF1_MmaPylRS PylRS M. mazei Mammalian TAA Simon Elsaesser...Mammalian TGA Simon Elsaesser 174901 pAS_MmaPylT_EF1_NES-MmaPylRS WT PylRS M. mazei Mammalian TAG Simon Elsaesser...

...Khakh 105869 pEMS1925 iCre-ERT2 Simpson 105870 pEMS1725 iCre-ERT2 Simpson 106368 pCMV-Tag2B-NCre N-terminal... iCre with MCS for inserting promoter none AAV Simpson 49455 pSH62-EBD Cre-EBD - Estradiol-dependent control...with MCS for inserting promoter, WPRE none AAV Simpson 72255 pCDH-CB-iCre-P2A-tdTomato-T2A-Puro iCre and...to turn off the expression of one gene while simultaneously turning on another. The FLEx switch system ...activates shRNA expression, removal of EGFP; See also similar plasmids pSico PGK GFP and pSico PGK puro Lentiviral...off shRNA expression, removal of EGFP; See also similar plasmids pSicoR PGK puro , pSicoR-Ef1a-mCh-Puro... to eGFP, removes dsRed. See article for other similar plasmids. Retroviral Clevers 15912 CTV Cre activates...

... every genomic target. Due to its comparative simplicity and adaptability, CRISPR rapidly became the most... change the genomic target of the Cas enzyme by simply changing the target sequence present in the gRNA...The requirements for a CRISPR knockout assay are simple. You’ll need a Cas enzyme and a gRNA specific to...AcrVA3.1, which inhibits both Cas12a and Cas3. Similarly, some Acr proteins are specific to a particular...editors are based on cytosine base editors and similarly work by first converting cytidine to uridine. ...repress gene expression in mice and human cells. The simplest dCas9-based activators and repressors consist ...imaging has numerous advantages, including the simplicity of gRNA design, programmability for different...

...assays that have multiple proteins to detect simultaneously. Conjugation The process of attaching signaling...conjugate or looking for antibodies validated for a similar assay type) and search again. After filtering, ..., directly compare the product information for similar antibodies, looking for ones that have identical...price, shipping availability, ease of purchase, or similar factors. Secondary antibodies Secondary antibodies...

...factor isoform has a unique 24 bp barcode that simplifies identification of the transcription factor isoform...is uniquely barcoded with a 24 bp sequence, to simplify identification during screening. Library ID PI...

...62201 Plant yes, cut S. pyogenes Hyg Chen pUC57-Simple-gRNA backbone 51306 Xenopus BsaI none S. pyogenes...Bacteria BsaI yes, activate S. pyogenes Marraffini pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 47868 Mammalian... natural type II-A CRISPR arrays capable of simultaneously repressing multiple target genes in Escherichia...

...sequences can be designed using Benchling or using similar design tools and ordered as standard desalted oligonucleotides...A., Park, K. U., & Brzezinski, J. A. (2020). Simultaneous deletion of Prdm1 and Vsx2 enhancers in the ...

...cleavage of the target. RNAi using siRNA follows a similar mechanism. Since it is pre-processed, it can bypass...., & Nemunaitis, J. (2009). siRNA vs. shRNA: Similarities and differences . Advanced Drug Delivery Reviews...

.... R., Abdul Aziz, A. T., Shueb, R. H., Zaid, M., Sim, B., Mustafa, M., Nik Yusof, N. K., Lee, C. K., Abu...

...Allows for visualization of multiple proteins simultaneously with dual labeling when used with two substrates...by ACP Synthase, thereby permitting selective simultaneous labeling in a sample. Please see the note below...

...target sites. (B) A TAL effector nuclease (TALEN). Similar in principle to the ZF nuclease, the components...with ZF arrays, if not slightly lower. Figure 2: Simplified representation of the Voytas/Bogdanove Golden...

...delivered to a host and generate virus (right), similar to how natural viruses can be delivered to a host...Viruses used to direct these alternate functions simply have to harbor alternate viral genomes, such that...

...Cas9 vector based on the Zhang lab's PX330 , simultaneously targeting the genomic site and the donor vector...Degron Tagging Masato Kanemaki's lab has developed a simple and scalable CRISPR/Cas-based method to tag endogenous...

...gene, as well as a target site common to a set of similar sequences. Organisms include human, mouse, rat...for NmCas9 and BsmBI site for cloning in gRNA: pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) PDF 47.5...