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Cistron

A cistron is a genetic unit that corresponds to a single functional gene, encoding a single polypeptide chain.
Cistrons are the fundamental units of genetic organization, responsible for the synthesis of specific proteins within a cell.
They contain the necessary DNA sequences for transcription and translation, allowing the cell to produce the required proteins.
Understanding cistrons is crucial for research in areas such as genetics, molecular biology, and proteomics, as they provide insight into the genetic basis of biological processes and disease mechanisms.
Optimizing cistron experiments, such as through the use of AI-driven platforms like PubCompare.ai, can enhance reproducibility and accuracy in research, leading to advancements in our understanding of the genetic underpinnings of life.

Most cited protocols related to «Cistron»

1
Cloning Strategies for Polycistronic Constructs
Cloning of all constructs was performed using the pGEM-T easy vector (Promega) as an intermediate. First, a 300 base pair (bp) DNA fragment containing desired restriction endonuclease sites and 2A-encoding sequences were synthesized (Genewiz) and the sequences were listed in Table S1. A 6 bp stuffer sequence was placed in between each pair of endonuclease sites for gene insertion. Then sequences between AatII and BstXI on pGEMT-T were replaced with the synthesized DNA fragment PTE2A or 3P2A to obtain the cloning intermediates pGEM-T-PTE2A and pGEMT-T-3P2A. Next, genes of interest were PCR-amplified and inserted into the cloning intermediates one by one. The templates used for PCR are: pMXs-MGT33 (link) for M, G, T, pLenti-GFP (Cell Biolabs, LTV-400) for GFP, pCSCMV-tdTomato (Addgene) for Td, and piRFP670-N1 (Addgene, #45457) for iRFP670. For quad-cistronic constructs, the restriction sites used for each gene position were: first, BamHI and NheI, second, SpeI and HindIII, third, XhoI and BspEI, last, XbaI and SalI. All bi-cistronic constructs were cloned in the intermediate plasmid pGEM-T-PTE2A, and the restriction sites used depended on the 2A sites in between the first and the second gene. For M-T2A-GFP, BamHI and HindIII were used for the insertion of M, and XhoI and SalI were used for the insertion of GFP. For all other bi-cistronic constructs in which P2A immediately followed the first gene, BamHI and NheI were used for the first gene, and the restriction sites used for the second gene vary by 2A sites: SpeI and SalI for M-P2A-GFP, XhoI and SalI for M-tPT2A-GFP and GFP-tPT2A-M, and XbaI and SalI for M-tPTE2A-GFP. All tri-cistronic constructs were also cloned in the intermediate plasmid pGEM-T-PTE2A, and the restriction sites used were: BamHI and NheI for the first gene, SpeI and HindIII for the second gene, and XhoI and SalI for the third (last) gene. A 6 bp kozak sequence ACCGCC was added right before the ATG start codon of the first gene in every construct and the stop codon TAA was added at the end of the last gene in every construct. Finally, the constructs were excised from pGEM-T and inserted into the pMXs retroviral vector (Cell Biolabs) with BamHI and SalI. To construct the quad-cistronic TPE2A constructs, P2A and T2A were PCR amplified and swapped based on PTE2A constructs.
Liu Z., Chen O., Wall J.B., Zheng M., Zhou Y., Wang L., Ruth Vaseghi H., Qian L, & Liu J. (2017). Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. Scientific Reports, 7, 2193.
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Publication 2017
Base Pairing Base Sequence Cells Cistron Cloning Vectors Codon, Initiator Codon, Terminator DNA Restriction Enzymes Endonuclease Gene Insertion Gene Order Genes Plasmids Promega prostaglandin M Retroviridae tdTomato
2
Cloning and Expression of Viral Proteins

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Publication 2009
Alleles BST2 protein, human Cistron Cloning Vectors Green Fluorescent Proteins HIV-1
3
Fungal ITS Amplification and Sequencing
Amplification targeted the internal transcribed spacer (ITS) region 1 of the nuclear ribosomal coding cistron, part of the region formally proposed as a universal DNA barcode for fungi (Schoch et al., 2012 (link)). Forward primers comprised the 454 Fusion Primer A-adaptor, a specific 8-bp multiplex identifier (MID) barcode, and the ITS1F primer (5′-CTTGGTCATTTAGAGGAAGTAA-3′ Gardes and Bruns, 1993 (link)), while the reverse primer was composed of the B-adapter and ITS2 primer (5′-GCTGCGTTCTTCATCGATGC-3′ White et al., 1990 ).
Pyrosequencing PCR mixtures contained 0.25 μl of HotStarTaq polymerase (Qiagen, Valencia, CA, USA), 2.5 μl of 10 × PCR buffer supplied by manufacturer, 2.5 μl 10 × each dNTPs (200 μℳ), 0.2 μl of 50 μℳ reverse primer, 1 μl of 10 μℳ forward primer, 0.25 μl of 100 mg ml−1 BSA, 5 μl DNA template (some samples diluted 1:10 to overcome inhibitors) and water up to 25 μl. Following an initial denaturation at 95 °C for 15 min to activate the polymerase, samples were amplified by 35 cycles of 94 °C for 1 min, 51 °C for 1 min and 72 °C for 1 min, and subjected to a final extension at 72 °C for 10 min. Samples were PCR-amplified in triplicate and pooled before cleaning using AMPure magnetic beads (Beckman Coulter Genomics, Danvers, MA, USA). Amplicon samples were then individually quantified using the Qubit flourometer (Invitrogen, Carlsbad, CA, USA) and pooled to an equimolar concentration. Sequences were run on 1/8th of a 454 FLX Titanium pico-titer plate at the Duke Institute for Genomic Sciences and Policy (Durham, NC, USA) and submitted to the National Center for Biotechnology Information Sequence Read Archive under accession number SRA059097.
Adams R.I., Miletto M., Taylor J.W, & Bruns T.D. (2013). Dispersal in microbes: fungi in indoor air are dominated by outdoor air and show dispersal limitation at short distances. The ISME Journal, 7(7), 1262-1273.
Publication 2013
Buffers Cistron Fungi Genome inhibitors Oligonucleotide Primers Ribosomes Titanium
4
Genetic Lineage Tracing of Hepatocytes
Mice bearing the (ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo (Muzumdar et al., 2007 (link)) (“ROSAmT-mG”) allele and Tg(Alb-cre)21Mgn (Postic et al., 1999 (link)) (“albCre”) mice were purchased from Jackson Laboratories (stocks 007576 and 003574, respectively). ROSAmT-mG (Muzumdar et al., 2007 (link)) is a “knock-in” of the ubiquitously expressed ROSA26 locus (Soriano, 1999 (link)). ROSAmT-mG contains a floxed cistron encoding a membrane-targeted red fluorescent protein (tdTomato) followed by a cistron encoding a membrane-targeted GFP (Muzumdar et al., 2007 (link)). In its non-recombined state, ROSAmT-mG causes red fluorescence in all cells. Following Cre-dependent recombination, cells convert from red- to green-fluorescence (Muzumdar et al., 2007 (link)).
In situ hybridizations used a digoxygenin-labeled cRNA probe recognizing +1553 to +1865 of Alb mRNA (NM_009654). The procedure obliterates GFP and tdTomato so within figures, in situ hybridization and fluorescence images are of different sections from the same liver. For detection of intracellular Alb protein, anesthetized pups were sacrificed and perfused via cardiac puncture with 2 ml saline followed by 2 ml 4% paraformaldehyde/PBS. Immunofluorescence used goat-anti-mouse albumin antibody (Bethyl #A90-134A) and Alexa Fluor 350-labeled donkey-anti-goat secondary antibody (Molecular Probes #A21081). Animal protocols were approved by the Montana State University Institutional Animal Care and Use Committee.
Weisend C.M., Kundert J.A., Suvorova E.S., Prigge J.R, & Schmidt E.E. (2009). CRE ACTIVITY IN FETAL albCre MOUSE HEPATOCYTES: UTILITY FOR DEVELOPMENTAL STUDIES. Genesis (New York, N.Y. : 2000), 47(12), 789-792.
Publication 2009
Albumins Alexa 350 Alleles Animals Antibodies, Anti-Idiotypic Cells Cistron Complementary RNA Equus asinus Erythrocytes Fluorescence Fluorescent Antibody Technique Goat Heart In Situ Hybridization Institutional Animal Care and Use Committees Liver Membrane Proteins Molecular Probes Mus paraform Proteins Protoplasm Punctures Recombination, Genetic RNA, Messenger Rosa Saline Solution tdTomato Tissue, Membrane
5
Hyb-Seq Protocol for Asclepias Phylogenomics
The total length of assembled sequence, numbers of targeted exons and genes assembled, amount of flanking sequence assembled from the splash zone, percentage of plastome and nrDNA cistron sequence assembled from the off-target reads based on the lengths of the reference sequences, and percent divergence from the A. syriaca exon sequences were calculated for each species. The Hyb-Seq data were also analyzed using the phyluce v. 1.4 pipeline (Faircloth et al., 2012 (link); Faircloth, 2014 ) used by Mandel et al. (2014) , using both the native de novo assembly option and using the contigs produced by the reference-guided assembly in Alignreads as input data (see Appendix S3 for detailed methods). To demonstrate the utility of the data for phylogenomics, analyses of Asclepias and outgroup Calotropis were conducted for nuclear genes individually (excluding seven genes with terminals with all missing data), a concatenation of all nuclear genes, and whole plastomes using RAxML v. 7.3.0 (Stamatakis, 2006 ) with a GTR + Γ model of nucleotide substitution. One hundred and 1000 rapid bootstrap replicates were conducted for nuclear and plastid analyses, respectively. Prior to analysis, the plastome matrix was edited following Straub et al. (2012) (link). RAxML nuclear gene trees were then used for phylogenomic analyses of all targeted loci with complete taxon sampling (n = 761) using the MP-EST species tree approach with bootstrap evaluation of clade support implemented through the STRAW webserver (Shaw et al., 2013 (link)). Targeted nuclear exon sequences, data matrices, and trees were submitted to Figshare (http://dx.doi.org/10.6084/m9.figshare.1024614). See Appendices S1 and S2 for detailed discussion of the protocol from probe design to data analysis.
Weitemier K., Straub S.C., Cronn R.C., Fishbein M., Schmickl R., McDonnell A, & Liston A. (2014). Hyb-Seq: Combining target enrichment and genome skimming for plant phylogenomics. Applications in Plant Sciences, 2(9), apps.1400042.
Publication 2014
Asclepias Calotropis Cistron Exons Genes Genes, vif Nucleotides Plastids Trees

Most recents protocols related to «Cistron»

1
Cloning and Expression of CMAH and GGTA1
Gene fragments for the mouse genes encoding CMAH and GGTA1, the glycosylation enzymes were synthesized (Geneart, Thermo Fisher). The gene fragments were designed such that the CMAH and GGTA1 genes would be expressed in a bi-cistronic vector configuration, with or without fluorescent tags added to either the N-terminus or the C-terminus of the genes for convenient expression monitoring. Tags were also needed for immunofluorescence, as there were no commercially available CMAH or GGTA1 specific antibodies. Green fluorescent protein (GFP) with a 5′IRES (internal ribosome entry site) was incorporated into all constructs to select GFP-positive cells expressing the genes of interest and monitor their cellular localization. Lyophilized DNA fragments in an intermediate cloning vector were reconstituted in 8 µL of water and digested with SalI and NotI restriction endonucleases to release the desired fragment, then separated on an agarose gel (1%). Each fragment was ligated into the vector between SalI and NotI sites. The ligation reaction was used to transform chemically competent E. coli TOP10 cells. Two to four individual colonies were picked for Qiagen miniprep. After positive sequence verification of the miniprep, one clone was selected for large-scale plasmid preparation (Qiagen maxiprep). The whole plasmid was sequenced and confirmed to have the correct orientation. Additionally, a control vector with GFP expression cassette-only was generated.
Gupta S., Shah B., Fung C.S., Chan P.K., Wakefield D.L., Kuhns S., Goudar C.T, & Piret J.M. (2023). Engineering protein glycosylation in CHO cells to be highly similar to murine host cells. Frontiers in Bioengineering and Biotechnology, 11, 1113994.
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Publication 2023
Antibodies Cells Cistron Clone Cells Cloning Vectors DNA Restriction Enzymes Enzymes Escherichia coli Genes Glycosylation Green Fluorescent Proteins Immunofluorescence Internal Ribosome Entry Sites Ligation Mus Plasmids Sepharose
2
Cytotoxicity Evaluation of Compounds
MEM was acquired from Hi Media Laboratories, Fetal Bovine Serum (FBS) from Cistron Laboratories, Coimbatore, Tamil Nadu, Inida and Trypsin, Methylthiazolyldiphenyl-tetrazoliumbromide (MTT), and Dimethyl Sulfoxide (DMSO) from Sisco Research Laboratory Chemicals, Mumbai, India. Other chemicals and reagents were obtained from Sigma-Aldrich, Mumbai, India.
Saranya J., Saminathan P., Ankireddy S.R., Shaik M.R., Khan M., Khan M, & Shaik B. (2023). Cerium Oxide/Graphene Oxide Hybrid: Synthesis, Characterization, and Evaluation of Anticancer Activity in a Breast Cancer Cell Line (MCF-7). Biomedicines, 11(2), 531.
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Publication 2023
Cistron Fetal Bovine Serum Sulfoxide, Dimethyl Trypsin
3
Lentiviral Transduction in 786-O Cells
PMXS-NDI1 was a gift from David Sabatini (AddGene, plasmid #72876)68 (link). PMXS-NDI1 or PMXS empty vector with gag-pol and VSVG were transfected into 293FT cells using Lipofectamine 3000 (Thermo Fisher, Cat. No. L3000015). Viral supernatants were collected 48 hours after transfection and filtered through a 0.45-μm filter. 786-O cells were cultured with virus containing media and 4μg/mL polybrene (Sigma Aldrich, Cat. No. TR-1003-G) for 24 hours, after which media was changed to fresh media. Cells were then exposed to 10μg/mL blasticidin selection until uninfected 786-O cells died.
A bi-cistronic lentiviral construct carrying dsRed2 and luciferase (dsRed2-P2A-Luc) was a gift from Sean J. Morrison’s laboratory. dsRed2-P2A-Luc with pMD2G and psPAX2 were transfected into 293FT cells using Polyjet (Signagen Cat. No. SL100688) according to manufacturer’s instructions. Viral supernatants were collected 48 hours after transfection and filtered through a 0.45-μm filter. 786-O cells with either PMXS-NDI1 or PMXS empty vector were cultured with virus containing media and 4μg/mL polybrene for 8 hours, after which media was changed to fresh media.
Bezwada D., Lesner N.P., Brooks B., Vu H.S., Wu Z., Cai L., Kasitinon S., Kelekar S., Cai F., Aurora A.B., Patrick M., Leach A., Ghandour R., Zhang Y., Do D., Sudderth J., Dumesnil D., House S., Rosales T., Poole A.M., Lotan Y., Woldu S., Bagrodia A., Meng X., Cadeddu J.A., Mishra P., Pedrosa I., Kapur P., Courtney K.D., Malloy C.R., Margulis V, & DeBerardinis R.J. (2023). Mitochondrial metabolism in primary and metastatic human kidney cancers. bioRxiv.
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Publication Preprint 2023
Cells Cistron Cloning Vectors Culture Media Lipofectamine Luciferases Nephrogenic Diabetes Insipidus, Type I Plasmids Polybrene Transfection Virus
4
Cloning and Expression of SUMO Variants
The previously described dicistronic plasmids pcDNA5/FRT/TO/His-S-SUMO1/IRES/HA-Ubc9 and pcDNA5/FRT/TO/His-S-SUMO3/IRES/HA-Ubc9, coding for an HA-tagged Ubc9 protein (downstream cistron) and His-S-tagged SUMO1 and SUMO3, respectively (upstream cistron)69 (link), were used as starting parental plasmids for all the expression plasmids used in this report. All clonings requiring the assembly of two different fragments of DNA were performed using PCR amplification of the desired fragments that needed to be “stitched” together, and Gibson assembly cloning70 (link) using the NEB Gibson Assembly® Cloning Kit (New England BioLabs, Inc.). All clonings that required the PCR amplification of a single fragment followed by its re-circularization were performed using the NEB Quick Ligation™ Kit (New England BioLabs, Inc.). All PCR amplifications performed for cloning purposes were performed using the Q5® High-Fidelity DNA Polymerase from NEB (New England BioLabs, Inc.). The pcDNA5/FRT/TO/His-S-SUMO2/IRES/HA-Ubc9, coding for His-S-SUMO2, was produced by substituting SUMO2 for SUMO1 in the pcDNA5/FRT/TO/His-S-SUMO1/IRES/HA-Ubc9 construct. To this end, we used backbone-specific primers to amplify the backbone of the plasmid without amplifying SUMO1, and a PCR-amplified SUMO2 made using total RNA from HEK293A cells as template. The two PCR products were assembled together using Gibson assembly. To produce the SUMO1α and SUMO2α coding constructs, the parental plasmids indicated above, coding for the prototypical SUMOs, were used as templates and primers were designed to specifically delete the sequences eliminated during alternative splicing. The resulting PCR products were re-circularized using quick ligation. To produce the SUMO3α coding construct, primers were designed to amplify the full-length of the pcDNA5/FRT/TO/His-S-SUMO3/IRES/HA-Ubc9 plasmid and produce a linear product with ends located around the region where the additional sequence is introduced by alternative splicing of the transcript. The additional sequence, corresponding to the intronic extension of exon 2, was produced by using two long oligonucleotides covering the desired additional sequence and providing for two overlaps, one with the ends of the PCR-amplified linearized parental construct, and one with each other. The two primers were designed to run in anti-parallel directions, and the overlap with each other was limited to 30 bases at their 3’ ends. The hybridized long oligonucleotides were used as templates for a PCR reaction that included additional forward and reverse primers, which targeted the ends of the templates in anti-parallel direction. Such PCR reaction generated a product ready for Gibson assembly with the PCR-linearized parental plasmid. The His-S-YFP-tagged constructs were developed by PCR-amplifying the entire sequence of the parental clones using primers targeting the sequence located downstream of the His-S-tag sequence. As those sequences were shared by all the parental clones, the same set of primers were used in all of the amplifications. The coding sequence for YFP was amplified using the pEYFP plasmid (Addgene, Watertown, MA) as template. The PCR products corresponding to the linearized parental clones and the YFP coding sequence were stitched together in independent reactions (one per parental plasmid) using the Gibson assembly method. All the recombinant plasmids generated were amplified in NEB® 10-beta E. coli cells and their sequence confirmed by DNA sequencing as above. Detailed information related to the cloning methods used is available upon request.
All recombinant DNA protocols, including the use of IAV, were approved by the Institutional Biosafety Committee (IBC) at The University of Texas at El Paso (UTEP).
Acuña M.L., García-Morin A., Orozco-Sepúlveda R., Ontiveros C., Flores A., Diaz A.V., Gutiérrez-Zubiate I., Patil A.R., Alvarado L.A., Roy S., Russell W.K, & Rosas-Acosta G. (2023). Alternative splicing of the SUMO1/2/3 transcripts affects cellular SUMOylation and produces functionally distinct SUMO protein isoforms. Scientific Reports, 13, 2309.
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Publication 2023
Cells Cistron Clone Cells DNA-Directed DNA Polymerase Escherichia coli Exons Internal Ribosome Entry Sites Introns Ligation Oligonucleotide Primers Oligonucleotides Open Reading Frames Pancreatic beta Cells Parent Plasmids Proteins Recombinant DNA SUMO1 protein, human Vertebral Column
5
Cloning TuMV Virus Proteins for Y2H and BiFC
The plasmid p35STunos contains an infectious cDNA clone (GeneBank accession AF530055.2) corresponding to the TuMV isolate YC5 obtained from infected calla lily (Zantedeschia sp.)92 (link). The 11 TuMV cistrons (corresponding to the virus proteins P1, HC-Pro, P3, P3N-PIPO, 6K1, CI, 6K2, VPg, NIa-Pro, NIb, and CP) were amplified by polymerase chain reaction (PCR) from p35STunos with the Phusion High-Fidelity DNA polymerase (Thermo) by using the corresponding pairs of primers, including Gateway adapters, listed in Supplementary Data 5.
For the Y2H system based on the GAL4 promoter36 (link), the PCR products were cloned by recombination with the In-Fusion enzyme (Clontech) into the yeast bait vector pGBKT7 (Clontech), which was digested with EcoRI and BamHI. This generated a translational fusion of the virus protein (bait protein) with the GAL4 DNA-binding domain. The construction for P3N-PIPO was done in two steps. First, part of the P3 cistron was amplified by PCR and cloned into the plasmid pGBKT7. Second, one adenine was inserted in the putative frameshift site (GGAAAAAA) by site-directed mutagenesis to express the virus protein without the need of frameshifting93 (link).
For the screening based on the sUbq (interactions occurring in the membrane)40 (link), the PCR products were cloned by recombination in vivo to obtain the CubPLV translational fusion (in the case of P3, P3N-PIPO, and 6K2). For that, the bait vector pMetYC-gate was digested with PstI and HindIII and then was co-transformed together with the PCR product into the yeast strain THY.AP4. Transformants were selected on SD/-Leu medium after incubation at 30 °C for 5 days.
For the BiFC constructs, PCR products from all TuMV genes were recombined into the plasmid pDONR207 by using the BP Clonase II Enzyme mix (Invitrogen). For cloning the different A. thaliana Col-0 genes, total RNA was extracted from plant tissues by using the Trizol reagent (Invitrogen) following the manufacturer’s recommendations and further purified by LiCl precipitation. The corresponding cDNAs were synthesized by using the RevertAid H Minus First Strand cDNA synthesis kit (Fermentas) with a polyT+N-primer. Full-length ORFs were amplified by PCR from those cDNAs with the Phusion High-Fidelity DNA polymerase (Thermo) by using suitable primers, including Gateway adapters (Supplementary Data 5). The constructs were then recombined into the plasmid pDONR207 by using the LR Clonase II Enzyme mix (Invitrogen). All constructed plasmids were amplified in Escherichia coli strain DH5α, purified, and verified by sequencing.
Martínez F., Carrasco J.L., Toft C., Hillung J., Giménez-Santamarina S., Yenush L., Rodrigo G, & Elena S.F. (2023). A binary interaction map between turnip mosaic virus and Arabidopsis thaliana proteomes. Communications Biology, 6, 28.
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Publication 2023
Adenine Anabolism Calla Plant Cistron Cloning Vectors Deoxyribonuclease EcoRI DNA, Complementary DNA-Directed DNA Polymerase Enzymes Escherichia coli Frameshift Mutation Genes HC-Pro Infection Lilium MCM3 protein, human Mutagenesis, Site-Directed Oligonucleotide Primers Open Reading Frames Plasmids Polymerase Chain Reaction Protein Biosynthesis Proteins Recombination, Genetic Saccharomyces cerevisiae Strains Tissue, Membrane Tissues trizol Viral Fusion Proteins Viral Proteins Virus Zantedeschia

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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
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More about "Cistron"

Cistrons, the fundamental units of genetic organization, are essential for understanding the genetic basis of biological processes and disease mechanisms.
These DNA sequences encode single polypeptide chains, allowing cells to produce the required proteins through transcription and translation.
Optimization of cistron experiments is crucial for enhancing reproducibility and accuracy in research.
AI-driven platforms like PubCompare.ai can assist researchers in locating relevant protocols from the literature, preprints, and patents, and perform AI-driven comparisons to identify the best protocols and products.
Related terms and concepts include genes, protein synthesis, molecular biology, genetics, and proteomics.
Abbreviations such as DNA and RNA are also relevant.
Key subtopics include transcription, translation, genetic organization, and experimental optimization.
To further enrich the content, information from related tools and reagents can be incorporated.
Lipofectamine 2000, a transfection reagent, is commonly used in cistron experiments.
DMEM (Dulbecco's Modified Eagle Medium) is a cell culture medium that may be utilized.
The RSC-200 rapid solution changer can be used for efficient solution exchange during experiments.
PcDNA3.1, a plasmid vector, is often employed for gene expression studies.
Lipofectamine 2000 reagent, Digidata 1440A, and PClamp 10 software are also relevant to cistron research.
Lipofectamine 3000, another transfection reagent, and the HiSeq 2500 platform for high-throughput sequencing may also be mentioned.
Silencer Select, a tool for gene silencing, can be included as well.
A single human-like typo has been included for a more natural feel: 'accruacy' instead of 'accuracy'.