Direct Mouse Genotyping Kit
The Direct Mouse Genotyping Kit is designed for rapid and convenient genomic DNA isolation and PCR amplification directly from mouse tissue samples. Typically used in mouse genetic screening and routine genotyping workflows, this kit employs optimized lysis and balancing buffer systems to release genomic DNA directly from tissue without conventional DNA purification steps. The provided lysate can be applied immediately as a PCR template, streamlining the overall experimental procedure. Moreover, the included ready-to-use 2X PCR Master Mix with dye facilitates PCR assay setup and ensures accurate amplification performance, making the kit suitable for high-throughput genetic analyses and routine applications in biomedical research labs.
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Related Biological Data
Related Biological Data
| 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. The Protease K solution should be stored at -20°C and could be stored at 4°C for a short time. Avoid repeated freeze/thaw cycles. Upon first use, it is recommended to aliquot sample into single use portions. | ||
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.
