Key Questions to Ask When Ordering qPCR Kits

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Bethyl Laboratories SARS-CoV-2 IgG ELISA Kit, www.bethyl.com

Bethyl Laboratories

Assays, kits, and reagents are used in many different life science, environmental, and research labs. Assay kits have become very important in providing the materials needed to conduct specific tests that require complex or hard-to-develop components like enzymes, antibodies, genomics, and cells. It is very important to purchase assays, kits, and reagents from trusted sources, as the experiments conducted will only be as good as the components in these products. For a list of laboratory assay, kit, and reagent manufacturers, see our online directory: LabManager.com/assays-kits-reagents-manufacturers

7 Questions to Ask When Buying Assays, Kits, and Reagents:

1. How pure do the components need to be for this application?
2. Is the same lot needed for an extended experiment?
3. How similar or dissimilar are the results from different lots?
4. Does the assay, kit, or reagent work well with the existing lab equipment?
5. Does the assay, kit, or reagent reduce lab work time to implement?
6. How important are potential contaminants from the packaging?
7. What is the unit price per test?

Quality Considerations

Given the wide range of assays, kits, and reagents available, and the large number of manufacturers, it is very important to understand the level of purity required, and the importance of trace components included. These can vary for items purchased from different vendors, and can even show important lot-to-lot variations from a single producer. Before starting a long-term set of experiments, it is worth understanding the precision required to obtain the desired results.

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If you are looking for more details, kindly visit TSKT.

Purchasing Tip

The recent pandemic has created havoc in some lab supply chains. Due to issues with supply and delivery of some assays, kits, and reagents, it is important to establish the robustness of the supply of important items for the lab. It may be necessary to have multiple sources of the key assays, kits, and reagents to ensure that the lab can continue to be productive, even in uncertain times.

When performing a RT it is possible to use three different primer types:

Oligo d(T) primers, which bind to the poly A tail of the RNA and then only transcribe RNA. This will avoid contamination with genomic DNA. As the poly A tail is located at the beginning of the gene it will also lead to more full transcripts.
Ramdom nonamers, which bind anywhere in the genome and allow the reverse transcriptase to fill up the gaps, will leads to high yields.
Specific primers, which bind to the gene of interest, and will therefore give specifics products.
The combination of oligo d(T) primers and random nonamers will give the highest yields and the longest transcripts, whereas specific primers transcribe only specific RNA but reduce the yield.
With an One step RT qPCR kit it is only possible to add specific primers, as it should be avoided that oligo d(T) primers and ramdom nonamers participate in the PCR reaction, giving many aspecific products. As a RT reaction is performed at 40-50°C, the primers can bind with mismatches to the RNA and therefore transcribe unwanted sequences, which then also will be amplified in to consecutive PCR, leading to aspecific PCR products. This disadvantage is inherent to the method.

In a Two step kit the oligo d(T) primers and the ramdom nonamers are included in the kit and will give ride to cDNA. Then two specific primers have to be selected and this will amplify the exact sequence.

If you want to learn more, please visit our website qPCR Kits.