Genotyping Kit
The Genotyping Kit for target alleles of insects, tissues, fishes and cells provides convenience in faster preparation and superior PCR amplification. Lysis buffer and balance buffer could digest tissues or cells rapidly to release unbroken genomic DNA, and it could be used as PCR template directly without extraction. There is no need for overnight digestion, Phenol/Chloroform extraction, toilsome manual purification or the utilization of expensive DNA spin columns. By using our kit, you can substantially shorten your time for digestion. Furthermore, the 2×PCR Master Mix (With Dye) will guarantee a robust and accurate amplification consequence. This Master Mix contains dye which means it can be directly electrophoresed after the amplification without the need to add a loading buffer. In addition, our genotyping Kit mitigates the risk of sample cross-contamination occurring in the PCR reaction by conducting the DNA extraction in one single tube.
| Complete Kit | 200 rxns | 500 rxns |
| Lysis buffer | 20 mL | 50 mL |
| Balance buffer | 20 mL | 50 mL |
| 2X PCR Master Mix (With Dye) | 1 mL x 2 | 1 mL x 5 |
| Proteinase K | 200 μL | 500 μL |
1. Lysis buffer and Balance buffer should be stored at 4°C. 2. 2X PCR Master Mix could be stored at -20°C for 2 years. 3. Store the unopened Protease K at -20 to -70 °C. Avoid repeated freeze/thaw cycles. Upon first use, it is recommended to aliquot sample into single use portions. The Proteinase K solution should be stored at -20°C and could be stored at 4°C for a short time. | ||
1. Q: What are the applications of the Genotyping Kits?
A: The Direct Mouse Genotyping Kit (Cat. No. K1025), Genotyping Kit (Cat. No. K1026), Direct Mouse Genotyping Kit Plus (Cat. No. K1027), and Direct Genotyping Kit Plus (Cat. No. K1504) can be used for genotyping mice, insects, fish, cells, and other samples. For example, you can use mouse tail, toes, ears, or other tissues for genotyping to quickly determine the genotype of the mice.
2. Q: What are the advantages of the Genotyping Kits?
A: The Master Mix in these kits contains loading dye, so the PCR products can be directly loaded for electrophoresis, making PCR more convenient. The lysis buffer and reaction buffer in the kits can rapidly digest tissues and release intact genomic DNA, without the need for overnight digestion, phenol/chloroform extraction, manual purification, or expensive DNA purification columns. The Genotyping Kits ensure stable and accurate amplification results.
3. Q: Why isn’t the tissue fully digested after lysis with the lysis buffer?
A: For most tissue samples, incubation with Proteinase K at 56°C for 15 minutes is sufficient to extract genomic DNA. The tissue may still appear intact, but lysis has already occurred, and the DNA obtained from partially digested tissue is usually adequate for PCR analysis.
4. Q: There are non-specific amplification bands when using the kit. What might be the reason?
A: Possible reasons include: improperly designed PCR primers; incorrect PCR reaction setup (e.g., DNA template concentration too high); inappropriate PCR cycling conditions (e.g., annealing temperature too low); or excessive ambient temperature during PCR mixture preparation.
5. Q: There is no target band in the PCR product. What could be the cause and how to solve it?
A: Possible causes and solutions include:
a) The tissue sample is not sufficiently digested; extend the digestion time.
b) Proteinase K is not fully inactivated; appropriately extend the incubation time at 95°C.
c) Primer quality issues; redesign the primers.
d) The target fragment has high GC content or is long; use the Direct Mouse Genotyping Kit Plus (Cat. No. K1027), which contains a genetically engineered Taq polymerase.
e) Inappropriate annealing temperature, insufficient extension time, or too few cycles; optimize the PCR conditions.
f) Template concentration is too low; increase the amount of template appropriately.
