|MPH02042||150 μl / 10 μM|
|Description||abm miRNA primers are for qPCR applications and each miRNA-specific primer set comes with a universal reverse primer. All miRNA primers are supplied in 10μM format.|
|Species||Human (H. sapiens)|
|Unit quantity||150 μl / 10 μM|
|Note||ABM miRNA All-In-One cDNA Synthesis Kit (Cat No. G898) must be used to reverse transcribe the RNA. Failure to do so will not yield valid qPCR results. Each primer set includes a universal reverse primer and is specific for the unique and proprietary sequence incorporated into the cDNA by the miRNA adapter in the miRNA cDNA synthesis kit. The miRNA qPCR primer sets cannot accurately detect cDNA generated using other first strand synthesis kits.|
|In what form are the miRNA primers sets supplied?|
We provide 150ul vials of each primer (1 x forward primer and 1 x universal reverse primer) at a concentration of 10uM.
|I want some information with regards to your quality methods of the primers to ensure that they are pure.|
Every oligo is deprotected and desalted to remove small molecule impurities. Oligos are quantified twice by UV spectrophotometry to provide an accurate measure of yield.
|What is the shipping temperature for the primers?|
The primers will be shipped with ice packs. If requested, room temperature for 1-2 days is fine but it is not recommended for optimal performance.
|What the Tm for the primer?|
We have validated all the primers with our own miRNA qPCR mastermix. For the qPCR set up instruction, please refer to page 6 of this file http://www.abmgood.com/miRNA/pdfs/miRNA%20HandbookV11.pdf Mastermixes from other companies may be used but the Tm of the primers will slightly vary. The Tm will need to be optimized.
|How does the universal reverse primer work?|
The universal reverse primer can be used for all mature miRNA. To quantify or amplify a specific miRNA, the universal reverse primer with the appropriate forward miRNA primer need to be used. Our forward primers can work with any kits. Our universal primers will only work with our cDNA synthesis kit. http://www.abmgood.com/miRNA-microRNA-cDNA-Synthesis-Kit.html
|How does the universal reverse primer work in amplifying mature miRNA?|
For PCR amplifying mature miRNA, total RNA needs to be isolated and reverse transcribed with our cDNA synthesis kit. A miRNA specific forward primer and an universal reverse primer (that recognizes a specific sequence incorporated to the cDNA) are used to amplify mature miRNA.
|How many tests can I perform with the given amount?|
Each primer unit is 10uM and 150ul. Thus, each unit will be approximately 250 qPCR reactions (final concentration of 300nM in 20ul final reaction volume).
|If we order primers (e.g. for miR126) Do you provide us with the sequence?|
We do not provide the sequences for our miRNA primers. However we do validate all of our primers in house and can verify their functionality.
|Are these primers compatible with the precursor form of miRNA?|
Our miRNA primer sets are compatible with the mature form only.
|I have ordered your miRNA primers and am using the Bio-Rad CFX Connect™ Real-Time PCR Detection System. Is your system a 1 color or 2 color system?|
Our EvaGreen miRNA qPCR Mastermixes is EvaGreen based and so the machine can only detect the EvaGreen/SYBR Green Channel. Please note that you will need to use abm's miRNA cDNA synthesis kit (Cat#G269/ G902) in order to use our primers/universal reverse primer.
- Sundar, I. K., Li, D., & Rahman, I. "Small RNA-sequence analysis of plasma-derived extracellular vesicle miRNAs in smokers and patients with chronic obstructive pulmonary disease as circulating biomarkers" Journal of Extracellular Vesicles 8(1):1684816 (2019).