Universal 3' miRNA Reverse Primer

Datasheet

  • Product Name

    Universal 3' miRNA Reverse Primer

  • Unit

    150 µl (10 µM) of each Forward and Reverse Primers

  • Description

    abm offers a comprehensive collection of miRNA qPCR primer sets which cover all known miRNA sequences.  This qPCR primer set consists of a unique Forward Primer specific for the miRNA and a Universal 3’ miRNA Reverse Primer which can be used for qPCR expression analysis. 

    The Universal 3' miRNA Reverse Primer is compatible with miRNA cDNA synthesized using abm’s miRNA All-In-One cDNA Synthesis Kit. This reverse primer is specific for the unique and proprietary sequence incorporated into the miRNA cDNA by the Oligo d(T) with Adapter in the miRNA All-In-One cDNA Synthesis Kit.  miRNA cDNA lacking this adapter sequence will not be compatible with this reverse primer.

  • System

    qPCR Primers

  • Storage Condition

    -20°C

  • Note

    Recommended Products for miRNA Expression Analysis:

    Product  Description 
    miRNA All-In-One cDNA Synthesis Kit abm’s miRNA qPCR primers must be used  with miRNA cDNA generated using abm’s miRNA All-In-One cDNA Synthesis Kit.

    BlasTaq™ qPCR MasterMix

    abm’s miRNA qPCR primers and miRNA cDNA generated from abm’s miRNA All-In-One cDNA Synthesis Kit can be combined with BlasTaq™ qPCR MasterMix for miRNA detection and quantification.

    Universal 3’ miRNA Reverse Primer

    This primer comes with each of abm’s miRNA qPCR Primer sets. It is also available separately for reorder.

    U6 Primers

    U6 is a small nuclear RNA (snRNA) that is sometimes used as a reference gene in order to normalize miRNA expression levels in certain cell and tissue samples.

    SNORD44 Primers

    SNORD44 is a small nucleolar RNA (snoRNA) that is often used as a reference gene in order to normalize miRNA expression levels.

    SNORD48 Primers

    SNORD48 is a small nucleolar RNA (snoRNA) that is often used as a reference gene in order to normalize miRNA expression levels.

    hsa-miR-16-5p Primers

    		hsa-miR-16-5p is a miRNA that is often used as a reference gene in order to normalize miRNA expression levels.
    Liao, G et al. "MicroRNA-203 negatively regulates c-Abl, ERK1/2 phosphorylation, and proliferation in smooth muscle cells" Physiol Rep 9:e12541 (2015). DOI: 10.14814/phy2.12541.
    Dawood, A. F., Younes, S., Alzamil, N. M., Alradini, F. A., & Saja, M. F. (2020). Inhibition of glycogen synthase kinase-3β protects against collagen type II-induced arthritis associated with the inhibition of miR155/24 and inflammation and upregulation of apoptosis in rats. Archives of Physiology and Biochemistry, 128(3), 679–687. https://doi.org/10.1080/13813455.2020.1716021
    Submit a review Submit a question
    Question
    Can the Universal Reverse miRNA primer work on any cDNA synthesis kit?
    ABM community
    Verified customer
    Asked on Jan 01 1970
    Answer
    The Universal Reverse miRNA primer can only work on our miRNA cDNA synthesis kit (cat#G269 and cat#G270). This is because our miRNA cDNA synthesis kit contains an unique dT adapter that allows the functionality of the universal reverse primer.
    ABM Scientific Support
    Answered on Jan 01 1970
    Question
    How should I store my lentivirus?
    ABM community
    Verified customer
    Asked on Mar 28 2025
    Answer
    The lentivirus should be aliquoted into smaller working volumes and then stored at -80°C. Lentivirus is sensitive to storage temperature and to freeze/thaws. It can lose up to 5% or more activity with each freeze/thaw. When stored properly, viral stocks of an appropriate titer should be suitable for use for up to one year. After long-term storage, we recommend re-titering your viral stocks before use.
    ABM Scientific Support
    Answered on Mar 28 2025
    Question
    Is it ready for use?
    ABM community
    Verified customer
    Asked on Jan 31 2023
    Answer
    Yes, it is ready to use. Just load suggested volume (5ul) to the well.
    ABM Scientific Support
    Answered on Jan 31 2023
    Question
    What generation does abm use for their lentiviral transfer vectors?
    ABM community
    Verified customer
    Asked on Feb 28 2025
    Answer
    abm lentiviral transfer vectors use the third generation lentivirus system. It is based on the inactivated HIV genome. Note that our lentivirus packaging plasmids cannot be integrated into the host and are transiently expressed.
    ABM Scientific Support
    Answered on Feb 28 2025
    Question
    Are abm’s lentiviral vectors compatible with my packaging mix?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    We recommend using abm’s 2nd Generation Packaging System Mix (Cat. No. LV003) or 3rd Generation Packaging System Mix (Cat. No. LV053). abm’s lentiviral vectors have been tested and are compatible with Invitrogen’s packaging mix; we have not tested other suppliers and cannot guarantee compatibility.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    Does MOI affect antibiotic resistance?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Higher MOI will provide more copies of the antibiotic resistance gene per cell. Cells containing multiple copies of the resistance gene can withstand higher antibiotic concentrations compared to those at lower MOIs. The concentration of antibiotic should be adjusted to a level that will cause selection for the desired population of transduced cells without going below the minimum antibiotic concentration you have established in your killing curve.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    How do I calculate the Multiplicity of Infection (MOI)?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    MOI (Multiplicity of Infection) refers to the number of viral particles per cell used in the infection, e.g. an MOI of 5 indicates that there are five infectious units (IU) or transducing units (TU) for every cell. MOI is determined by calculating the numbers of viral particles added per well then divide this number by the number of cells seeded into the well. We also recommend transducing the cells with a range of MOIs as different cell types may require different MOIs for successful transduction.

    MOI = Product Titer (IU/ml) x Virus Volume (ml) / Total Cell Number
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    What concentration of puromycin should I use for cell selection following lentivirus transduction?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    The concentration must be optimized for each cell type.  Typical selection amounts are around 0.1 - 0.5µg per ml.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    Are abm’s lentiviral vectors compatible with packaging plamids psPAX2 and pDM2.G?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Yes.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    What is the optimal cell density for lentivirus transduction?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Optimal cell density for lentiviral transduction is generally 50-70% confluent for most cell types. Certain cell lines may require the optimal cell density to be determined experimentally.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    Are abm’s vectors high or low copy? What is the expected yield from a plasmid extraction prep?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    These are medium-high copy plasmids and should be propagated in a cloning E. coli strain such as DH5α. Typical yields from a 250ml culture is 300-500µg plasmid DNA.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    What kind of cDNA is used for these primers? How do I prepare the cDNA?
    ABM community
    Verified customer
    Asked on Jan 01 1970
    Answer
    cDNA can be prepared from our cDNA synthesis kits, further information can be found from our miRNA page; http://www.abmgood.com/miRNA-microRNA.html
    ABM Scientific Support
    Answered on Jan 01 1970
    Question
    Do abm’s lentiviral vectors contain a chimeric RSV promoter in the upstream 5’ LTR?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Yes.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    How are abm’s vectors shipped and what buffer are they supplied in?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Standard delivery of abm’s vectors is supplied in TE buffer (10mM Tris, 1mM EDTA, pH 8.0). Our vectors can also be ordered and delivered in agar stabs for an additional fee.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    Do you recommend using 2nd or 3rd generation lentivirus packaging mix?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    abm's lentiviral vectors are compatible with 2nd and 3rd generation packaging systems. We recommend abm’s Second Generation Packaging Mix (Cat. No. LV003) for users who want to achieve higher titer lentivirus as only 3 plasmids are required for virus production instead of 4 plasmids.  We also recommend abm’s Third Generation Packaging Mix (Cat. No. LV053) for users concerned with virus biosafety, as the viral packaging elements are further split into 4 plasmids thus reducing probably of producing wild type virus.

    Please note, only packaging mixes produced by abm have been tested in house and therefore carry our guarantee for high titer virus production. If it is desirable to use other packaging plasmids obtained from a different source, the compatibility must be tested and determined by the end user.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    Will these primers work for any species?
    ABM community
    Verified customer
    Asked on Jan 01 1970
    Answer
    Our Universal 3' miRNA Reverse Primers (Cat# MPH00000) should work for any species including human and mice. Please note that our universal 3' miRNA Reverse Primers will only work with abm's miRNA cDNA synthesis kits (cat#G269/G270/G902/G903).
    ABM Scientific Support
    Answered on Jan 01 1970
    Question
    Can I insert my own polyA signal into abm’s lentiviral vector? Where is the polyA signal located in abm’s lentiviral vectors?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Since lentivirus is an RNA virus, during the synthesis of the RNA genome to be packaged, if there is a polyadenylation signal in the insert, the RNA will be terminated prematurely. There is a SV40 poly(A) signal in the vector located downstream of the 3'LTR. It is designed this way to ensure a high amount of transcriptional RNA is present so that a high viral titer is obtained. We have received positive feedback from previous customers in regards to the high and stable expression levels of our lentivectors.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    What concentration are abm’s vectors supplied as?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Typically vectors are supplied at 100ng/µl.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    Do abm’s lentiviral vectors contain the WPRE?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    Yes, all of abm’s lentiviral vectors contain the WPRE (woodchuck hepatitis virus post-transcriptional regulatory element) upstream of the 3’LTR. This element when transcribed creates a tertiary structure which enhances gene expression.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    How should I store abm’s vectors?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    We recommend storing abm’s vectors at -20°C and avoiding repeated freeze/thaws as this may affect plasmid integrity and cloning efficiency.
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    What is MOI and how do I know which MOI to use?
    ABM community
    Verified customer
    Asked on Mar 24 2025
    Answer
    MOI stands for multiplicity of infection. Theoretically, an MOI of 1 will provide 1 virus particle for each cell on a plate, while an MOI of 10 represents ten virus particles per cell. However, several factors can influence the optimal MOI including cell line, cell type, transduction efficiency and gene of interest. We recommend first establishing an optimal MOI for each cell line. This can be done using a range of MOIs (0, 0.5, 1, 2, 5, 10, 50) to determine the MOI required to obtain optimal gene expression
    ABM Scientific Support
    Answered on Mar 24 2025
    Question
    What buffer is abm’s lentivirus provided in?
    ABM community
    Verified customer
    Asked on Mar 28 2025
    Answer
    10^7 IU/ml = DMEM
    10^8, 10^9, 10^10 IU/ml = 1X PBS (pH 7.4) with 2% PEG6000
    ABM Scientific Support
    Answered on Mar 28 2025
    Question
    How do I transform abm’s plasmids?
    ABM community
    Verified customer
    Asked on Mar 25 2025
    Answer
    We recommend transforming abm’s plasmids into our ProClone™ Competent Cells (Cat. No. E003) and following the protocol outlined in the technical Datasheet. The ProClone™ Competent Cells are high-efficiency chemically competent DH5α (E. coli) cells which are ideal for plasmid amplification.
    ABM Scientific Support
    Answered on Mar 25 2025
    Question
    How do I revive an agar stab?
    ABM community
    Verified customer
    Asked on Mar 25 2025
    Answer
    1. Use a sterile loop or pipette tip to scrape a small amount of cells from the agar stab. Only a tiny amount is needed as it contains live E. coli.
    2. Gently streak the cells onto an agar plate containing the correct antibiotic selection in order to achieve single isolated colonies.
    3. Place the plate “agar side up” and incubate at 37°C overnight.
    4. Select a single colony from the plate for downstream applications, such as an overnight broth culture and grow E. coli to late exponential-early stationary phase. 4a. The broth culture can be subjected to miniprep plasmid extraction. We recommend using abm’s Column-Pure Plasmid Miniprep Kit (Cat. No. G4003). 4b. The broth culture can be used to prepare a glycerol stock for long-term storage. Mix culture with sterile glycerol to a final concentration of 15% and store at -80°C.
    ABM Scientific Support
    Answered on Mar 25 2025
    Question
    I packaged abm’s lentivector into virus, performed transduction and antibiotic selection. Following selection all my cells died. What should I do?
    ABM community
    Verified customer
    Asked on Mar 25 2025
    Answer
    We recommend first checking the following parameters:
    1. Cell density. If density is too low, this can also result in cell death.
    We recommend the following experiments:
    Initial Experiment
    •Transduce lentivirus into the 293T cell line - one of the easiest cell lines to transduce. If the transduced cells survive selection, then troubleshooting should focus on post-virus production steps. If the transduced cells didn't survive selection, the troubleshooting should focus on virus packaging steps.
    Virus Packaging Troubleshooting
    •Attempt lentivirus packaging using a GFP expressing control vector. If packaging is successful with the control, then repeat packing of the lentivirus in question with a GFP batch control to ensure no errors with the first production. If no GFP signal is observed after transfection or ~72h after a test transduction of a GFP control lentivirus on 293T cells, then the packaging process needs to be assessed.  Packaging process assessment includes reagent quality, cell health, and packaging protocol details.
    Post-Virus Production Troubleshooting
    •Assess if the correct antibiotic concentration is used. This can be done by performing a drug-killing curve. To set up a drug-killing curve, we recommend using the same culture size and seeding density for your actual selection, and adding a different puromycin concentration to each sample, with the range between 0-1 ug/ml. If the cells are not killed at the 1ug/ml, you may try increasing the range higher concentrations. It is important to identify the concentration that results in >95% cell death in 1-4 days to establish the minimal concentration to use for the selection process.
    •Assess if the correct MOI is used to transduce the target cells. This can be done by using a GFP control lentivirus to transduce the target cells at a range around literature-recommended MOIs to determine the optimal MOI for transduction.
    •Assess if a transduction enhancer such as the ViralEntry Transduction Enhancer (Cat.No. G515) is necessary. Some cell lines are more difficult to transduce than others and by using a transduction enhancer can lead to increased target cell permeability.
    ABM Scientific Support
    Answered on Mar 25 2025
    Question
    I transduced my cell line using abm’s virus. Afterwards my cells exhibited a different morphology and/or died. Why?
    ABM community
    Verified customer
    Asked on Mar 25 2025
    Answer
    1. This often happens to primary cells which are very sensitive to culture medium conditions. Try to use a higher titer virus in order to limit the volume of exogenous media added to your cells.
    2. Virus preparation might be contaminated with bacteria. Use a 0.25µm syringe filter to clear the virus preparation and repeat transduction following strict sterile technique.
    3. Cell line might be contaminated with mycoplasm. This type of contamination is often not immediately obvious. The contamination effect is amplified after virus transduction. Repeat transduction with fresh cells.
    4. The virus preparation may contain some cell debris which could be mistaken as a change in cell morphology. Normally this issue will disappear 3-5 days after virus transduction.
    ABM Scientific Support
    Answered on Mar 25 2025
    Question
    I am interested in one of abm’s vectors. Where can I find the corresponding control?
    ABM community
    Verified customer
    Asked on Mar 25 2025
    Answer
    A complete list of controls can be found on abm’s Control Vectors and Viruses page. https://www.abmgood.com/Control-Vectors-and-Viruses.html
    ABM Scientific Support
    Answered on Mar 25 2025
    Question
    I transduced abm’s lentivirus into my target cell line but there is no fluorescence or very weak signal. Why?
    ABM community
    Verified customer
    Asked on Mar 25 2025
    Answer
    There could many possible reasons for this result.
    1. Typically GFP is behind a weaker promoter such as SV40 or CBH leading to a less robust fluorescence signal.
    2. Virus titer is low leading to only a small proportion of successfully transduced cells. Try increasing the MOI or using a transduction enhancer such as abm’s ViralEntry™ (Cat.No. G516). 3. Target cells are difficult to transduce. A higher MOI must be tested. Some cells are very easy to be transduced such as 293T, while others such as lymphocytes are very difficult.
    4. Lentivirus became inactivated during transportation and storage. Lentivirus is very sensitive to temperature changes. Leaving lentivirus at room temperature, 4°C for 48 hours, or subjecting it to multiple freeze/thaws can lead to inactivation or reduced titer.
    ABM Scientific Support
    Answered on Mar 25 2025
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Current vector selected:
Universal 3' miRNA Reverse Primer
Cat. No.
MPH00000
Plasmid DNA Services
Midiprep Plasmid Amplification
10-20 µg
$75.00
Maxiprep Plasmid Amplification
80-100 µg
$150.00
Bacterial Agar Stab
1 stab
$40.00
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Lenti-III-mir-GFP Control Virus
m002
4 x 500 µl
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ViralEntry™ Transduction Enhancer (100X)
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$190.00
pLenti-III-mir-Off Control Vector
m007
500 ng
$93.00
Lenti-III-mir-Off Control Virus
m008
4 x 500 µl
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Current vector selected:
Cat. No.
Controls and Related Products
miRNA All-In-One cDNA Synthesis Kit
G898
20 rxn
$185.00
pLenti-III-mir-GFP Control Vector
m001
500 ng
$93.00
Lenti-III-mir-GFP Control Virus
m002
4 x 500 µl
$212.00
ViralEntry™ Transduction Enhancer (100X)
G515
1.0 ml
$190.00
pLenti-III-mir-Off Control Vector
m007
500 ng
$93.00
Lenti-III-mir-Off Control Virus
m008
4 x 500 µl
$212.00
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