GeneCopoeia offers comprehensive qRT-PCR tools, including first-strand cDNA synthesis kits, qPCR mix, RT-PCR detection kits, validated qPCR primers and qPCR arrays for researchers to study gene and microRNA (miRNA) expressions.
96-well or 384-well qPCR arrays containing pre-defined or customized groups of qPCR primers (fully validated) to detect expression profiles of genes or miRNAs in various tissues and cells
The Small Ubiquitin MOdifier (SUMO) gene fusion system allows for the efficient removal of SUMO tags. The CoolCutter™ SUMO Protease product is a mixture of recombinant human and mouse SUMO proteases which delivers superior SUMO cleavage activity with both the native sumoylated proteins and the SUMO tag in recombinant SUMO fusion proteins. The CoolCutter™ enzyme recognizes the tertiary structure of SUMO rather than an amino acid sequence for a clean release of the desired peptides. A high activity in a wide range of pH, salt and detergent concentrations makes CoolCutter™ SUMO protease highly suitable for recombinant protein expression and purification. CoolCutter™ SUMO protease can cleave any protein that is fused to the C-terminus of SUMO except for fused proteins beginning with proline.Cleavage efficiency of CoolCutter SUMO Protease is low for those SUMO-fused proteins that begin with luecine, lysine or valine. CoolCutter™ SUMO Protease is ideal for use with OmicsLink™ Expression-Ready Clones, particularly the bacterial pReceiver-B12 and pReceiver-B13 vectors.
CoolCutter™ SUMO Protease and no-gap cloning
CoolCutter™ SUMO protease allows construction of full-length ORF cDNA clones that produce proteins without spacer peptides between the tag and desired proteins.
Other proteases contain cleavage sites resulting in unwanted spacer peptides
CoolCutter™ SUMO protease generates full- length protein without spacer peptides
Figure 1. Western blot showing cleavage efficiency of CoolCutter™ SUMO protease at 16°C and 25°C for 3-, 5-, 10-, 20- and 40-minute incubations.
Description: The single strand cDNA synthesis kit is designed for the purpose of producing first strand cDNA using polyA RNA(or total RNA) as templates. The synthesized cDNA can be used as templates in PCR to detect gene expression. It is a complementary product of the AmpArray product. The first strand cDNA can also be used to generate the gene fragment for gene cloning.
The components included in the kit:
5x M-MLV reaction buffere
200µl
Oligo dT(0.5 µg/µl)
20µl
dNTP mix (2.5 mM each)
200µl
M-MLV reverse transcriptase
5,000/25µl
Protocol: The protocol for the kit will be shipped together with all components
Price:
Cat. No.
First Strand cDNA synthesis kit (10 reaction)
Qty
Price($USD)
SSSK-01
Including following componants:
1. 5x first strand buffer
2. Oligo dT
3. dNTP mix
4. Reverse Transcriptase MMLV
GeneCopoeia offers genome-wide human, mouse and rat microRNA (miRNA) 3′ UTR target clones in mammalian expression vectors. miRNA 3′ UTR target clones can be used for miRNA target identification and functional validation of predicted targets, or to study the regulatory effect of miRNAs on target genes.
3′ UTR sequences were obtained from public domain gene sequence databases and inserted downstream of a secreted Gaussia luciferase (GLuc) reporter gene, which is expressed under control of the SV40 promoter in mammalian cells. A chimeric mRNA is transcribed consisting of the GLuc and a 3′ UTR target sequence. Thus, the study of mRNA-miRNA target interaction can be easily performed with a live cell assay for GLuc using only 10 μl of the cell culture medium.
Besides using GLuc as the miRNA 3′ UTR target reporter, a secreted Alkaline Phosphatase (SEAP) reporter driven by a CMV promoter is also cloned into the same vector (pEZX-MT05) and serves as the internal control. The dual-reporter vector system enables transfection normalization for accurate across-sample comparison.
In addition to the GLuc/SEAP dual-reporter vector system, a firefly/Renilla Duo-Luciferase reporter vector (pEZX-MT06) is also available.
Figure 3. The inhibitory effect of miRNA on a 3′ UTR target sequence expression clone
LIN28 is the known target gene for miR-125a. HEK 293 cells were transfected with LIN28 3′ UTR target sequence expression plasmid alone (left bar) or co-transfected with miR-125a precursor expression plasmid (right bar). Both the GLuc activity and an internal control SEAP activity were determined 24 hours post-transfection. The activity ratio of GLuc to SEAP was set to 1 for the single transfection sample with LIN28 3′-UTR target sequence expression clone (left bar), against which the activity of the co-transfection sample was compared to. The result shows that miR-125a suppressed the luciferase activity from the GLuc-LIN28-3′-UTR clone by more than 70% (right bar).
Advantages
Live cell assays
Naturally secreted GLuc reporter
No lysis of the cells is necessary
Save samples, reduce variations, and simplify experiments
Dual-reporter vector system
Enables transfection normalization for accurate across-sample comparison
High-throughput compatible
Group study or high sample number compatible
High sensitivity
GLuc is 1000-fold more sensitive than firefly or Renilla luciferase
*Promotions are valid in the US & Canada only. For international customers, please contact your local distributors. Discounts are not valid on previous purchases and cannot be combined with additional discounts.
MicroRNAs (miRNAs) are small, non-coding, single-stranded RNA molecules found in eukaryotic organisms. They are highly conserved and usually 21-23 nucleotides in length. miRNAs are involved in almost all cellular functions and regulate gene expression by binding to the 3′ untranslated regions (3′ UTRs) of targeted mRNAs. Irregularities in miRNA-regulated gene expression have been found to be associated with cancers, cardiovascular disorders and a variety of other diseases.
GeneCopoeia offers miExpress™ precursor miRNA expression clones in lentiviral and non-viral vectors with eGFP reporter gene. Hairpin precursor miRNA of approximately 150 nucleotides is cloned into lentiviral or non-viral vectors for delivery in virtually all cell types. Clone sets are also available.
Search precursor miRNA clones and synthetic miRNA mimics
Advantages
Full coverage
All known human, mouse and rat miRNAs in the current miRBase covered.
Flexible delivery systems
Lentiviral vectors allow efficient transduction and stable integration into the host genome of non-dividing and difficult to transfect cells target cells
Non-viral vectors allow either transient or stable transfection
Optimized expression cassettes
Expression cassettes have been fully sequenced and optimized for high miRNA expression of precursor miRNA and maturation of miRNA inside cells
Selection markers and reporters
An eGFP reporter is designed to monitor transfection or transduction efficiency
Neomycin or Puromycin selection marker enables stable cell line selection
Figure 2: Role of precursor miRNA and miRNA 3′ UTR target clones in miRNA regulation studies
Besides individual clones, GeneCopoeia also offers pre-made or custom-made miRNA precursor clone sets. Uniformity and high-quality clone sets make them ideal for large-scale high content and high-throughput screening studies in functional genomics, proteomics and systems biology.
Clone sets by gene family in the vector of your choice
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