For many reasons, genotyping has settled into our everyday lives. It provides information for our medical records, scientific research and even for antibiotic resistance.
But first, what is Genotyping?
Genotyping is a technology that determines an individual or animal’s genotype. Genotypes are usually small genetic differences which could cause important physical changes. For example, in Albinism, a mutated TYR gene can cause the non-synthesis of melanin. This leads to the characteristic pale skin, white hair and light-coloured eyes. Genotyping can be used to find the differences in a DNA sequence by comparing it to another sample. It works by identifying the variations in the DNA sequence. This is particularly useful for understanding genetic diseases like Huntington’s disease and Sickle cell anaemia—since most of them are caused by a mutation or variation in the DNA sequences.
Genotyping is also used in scientific research to find out the genotype of mice or other animals used in a study. This information provides answers to certain physical traits the animal might possess or lack. A popular and fast method of determining genotype is through PCR or Polymerase Chain Reaction genotyping.
How does PCR Genotyping work?
PCR genotyping kits contain both lysis solution and master mix for the PCR. Since the PCR depends on having DNA to examine, you have to release your DNA of interest from the cell first. You can do this by adding the tissue (where you want to extract your DNA) or mouse-ear punch into the lysing solution. Thereafter perform PCR using the master mix (if you are using the Microzone Genotyping kit). However, this method may vary depending on your laboratory protocol.
However, PCR genotyping is a sensitive test and should be performed carefully. In some cases, you may get irregular or wrong results. Here are 3 mistakes you should avoid when performing PCR genotyping.
3 Mistakes You Should Avoid In PCR Genotyping
1. Contamination
You should ensure that all forceps and tubes for handling the samples are clean in between animals. A drop of blood or hair from one sample on another causes contamination. This often leads to inaccurate results.
2. Using more than the stipulated time
Some operations in PCR genotyping usually have a stipulated time for them to run. For example, when using our Clent Life Genotyping kit, Vortex time is five minutes for cell lysis. Taking more time than necessary to vortex can lead to mechanical shearing of the DNA or a breakdown of the DNA into small pieces. This makes it harder to isolate the DNA.
3. Preserving samples wrongly
Several reagents used during PCR must be preserved properly if not in use. Whilst carrying out the PCR genotyping, you may not use up all the master mix for PCR. In this case, you can refreeze the unused master mix (which contains DNA primers) to preserve it. However, you should ensure that you refreeze them properly to avoid errors when you wish to use them.
Our PCR Genotyping Kit
Our PCR Genotyping kit comes with a lysis solution and Megamix for your PCR Genotyping. You can go from sample to gel with speed and efficiency.
For DNA Extraction, our fast, easy, single tube lysis helps you get quality PCR-ready DNA in just 15 minutes.
For PCR amplification, Microzone’s 2x MegaMix-GT PCR master mix includes an enhanced PCR reaction buffer, dNTPs and an inert blue loading dye to enable direct gel loading post PCR.
Conclusion
Although PCR genotyping is a sensitive test, it is also a fast, accurate and efficient method of determining genotype.
To learn more about our genotyping kits, visit here.