BlasTaq™ 2X qPCR MasterMix
|G892||2,500 rxn (25.0 ml)|
BlasTaq™ 2X qPCR MasterMix provides a convenient, reliable and robust setup for performing quantitative real-time analysis of DNA samples. This ready-to-use qPCR MasterMix contains abm’s strategically-engineered, next generation Taq Polymerase, BlasTaqTM DNA Polymerase, providing for rapid extension rates and robust performance. With specialized reaction conditions, this polymerase provides increased processivity, yields, and sensitivity, while shortening reaction times by up to 70%, compared to wild-type Taq DNA polymerase.
BlasTaq™ has 5’-3’ polymerase and 5’-3’ exonuclease activities, lacks 3’-5’ exonuclease activity, and produces 3’-dA-tailed amplicons. qPCR products made with BlasTaq™ can be used with TA cloning vectors.
BlasTaq™ 2X qPCR MasterMix comes with a separate vial of ROX Reference Dye which can be added depending on the qPCR machine type, as listed in the table below.
Store at -20°C.
|Unit quantity||2,500 rxn (25.0 ml)|
|Which species are covered by a miRNA qPCR array?|
The complete layout and miRNA targets included in our arrays can be found on the product page. Currently, ABM offers miRNA qPCR arrays for whole human and mouse genomes, as annotated by miRBase Release 16.0.
|I still have additional questions, who do I contact?|
For further details, please email [email protected] or 1-866-757-2414.
|Can ABM provide assistance with summarizing the miRNA qPCR array for my publications?|
Sure, please contact us for publication assistance.
|Where can I view performance data?|
1nt performance data for our high performance arrays can be found on our general qPCR miRNA Array page.
|Does ABM provide a miRNA profiling service?|
Yes, we provide a miRNA profiling service using our latest generation of miRNA arrays. Visit our miRNA custom services FAQs or the miRNA profiling services page for more information.
|What type of RNA internal standards are there for quantitative determination of miRNA?|
Our arrays include 4 endogenous controls for normalization of your qPCR data. The human endogenous controls are snoRD44, snoRD47, snoRD48, and U6-2. For mouse, the endogenous controls are Rt U1, U6, RNU43, and snoRNA142. All our endogenous controls have been validated using multiple sources of RNA.
|Which qPCR machine do I need to read miRNA arrays?|
You may use any 384-well qPCR machine, but remember to specify your machine in your order so your array contains the right master mix for your machine.
|How many miRNA species are included in the arrays?|
Our Human and Mouse Whole Genome miRNA qPCR arrays cover 1034 and 726 miRNA mature species, respectively.
|What reagents do I need to perform a qPCR miRNA array experiment?|
All the reagents you will require for the qPCR miRNA profiling are contained within the array you order from ABM. The kit includes 5ml of complimentary 2X BrightGreen qPCR miRNA mastermix, sufficient for two 384-arrays. Additional mastermix can be ordered online or by contacting [email protected]
|What is the sequence of the universal primer contained in the miRNA qPCR arrays?|
The sequence of the universal primer is proprietary, and unfortunately cannot be disclosed under any circumstances.
|What is in the wells of your qPCR arrays?|
Each well contains 4ul of primer mixture, including miRNA specific and universal primer. Our optimized qPCR Mastermix is provided in a separate tube.
|How many biological replicates do I need for each condition?|
To increase confidence and reduce experimental error, it is suggested that you use at least 3 replicates per sample. Alternatively, you may wish to use only one sample per condition; pick out miRNA of interest using our analysis spreadsheet (or do your own analysis); and then use replicates for select miRNA species. You may contact a representative at [email protected] for quotes and details on individual miRNA assays. Or, order individual miRNA primer sets online.
|Can precursor miRNAs be detected on the qPCR array?|
Each primer set has been designed and tested to detect only mature miRNA.
|Is there a way I can check for the presence of small RNA in my total RNA sample before analyzing?|
Yes, this can easily be done with a 1-2% agarose gel. You should see a 4.5kb band at double the intensity of a 1.9kb band; as well you should not see any smearing, as this is a sign of RNA degradation. If you do not have gel documentation equipment, a similar analysis can be done using a UV spectrometer. Ensure the 260nm/280nm ratio is 1.8, while the 260nm/230nm ratio is >1.0. A 260nm/280nm ratio >2.0 indicates unwanted residual ethanol while a ratio <1.5 suggests RNA degradation or sample contamination.
|Do I need to enrich my total RNA sample for small RNAs?|
No, our miRNA profiling system is designed and optimized to detect all miRNA species starting with total RNA.
|How do I prepare my total RNA sample?|
Trizol reagent is recommended. You may also use other methods as long as it preserves the total RNA including small size RNA within your sample and minimizes the amount of solvent left in the sample.
|Is it possible to use your kit (miRNA cDNA Synthesis Kit) to do quantitative PCR with the Taqman strategy?|
This kit along with our miRNA primers would not be compatible for use with Taqman strategy.
|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 needs to be used. Our forward primers can work with any kits. Our universal primers will only work with abm's cDNA synthesis kit.
|Can I use SYBR green mastermix for downstream qPCR of the cDNA produced with this kit?|
It may be possible to use a SYBR green mastermix for qPCR, however, our BrightGreen miRNA miRNA qPCR MasterMix recipe has been fully optimized to provide the most successful conditions for qPCR. We have performed extensive testing with minor differences in the composition of miRNA qPCR buffer, this has proven to give very dramatic differences to the results of qPCR. General qPCR mastermixes have more additives to prevent non-specificity and formation of primer dimers and the conditions are often too harsh for the miRNA primers to anneal properly
|Is the dye set-up similar to SYBR Green™ & EvaGreen™?|
Yes, it is comparable.
|What are the advantages to using BlasTaq™ 2X qPCR MasterMix?|
It is a ready-to-use qPCR MasterMix in which only template and primers need to be added by the end user. BlasTaq™ has super speed performance to achieve results less than a hour. ROX Reference Dye is offered on the side to give more flexibility to the user for your qPCR set up.
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