Direct Genotyping Kit Plus
The Direct Genotyping Kit Plus is specifically designed for species genotype identification. This product contains lysis buffer and balance buffer, which can quickly digest tissues, release complete genomic DNA, and the DNA can be directly used as a PCR template without being extracted from the mixed solution. The kit contains modified Taq DNA Polymerase, dNTP and an optimized buffer system. The modified Taq DNA Polymerase has strong amplification performance and is compatible with a variety of complex templates. It is suitable for gene identification of various tissues or cells including plant, colony PCR and mice. This Master Mix operation only requires the addition of primers, templates and ddH2O, which greatly simplifies the experimental steps, reduces personal errors and improves the repeatability of the results. This product contains blue dye, which means it can be directly subjected to electrophoresis after amplification without the need to add sample loading buffer. If agarose Gel electrophoresis of PCR products is required, our product SYBR Safe DNA Gel Stain (Item No. A8743) can be added to the gel.
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Components |
200 rxns |
500 rxns |
Storage |
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2× Taq PCR Master Mix Plus (with dye) |
2×1 mL |
5×1 mL |
-20°C |
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Balance buffer |
20 mL |
50 mL |
4°C |
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Lysis buffer |
20 mL |
50 mL |
4°C |
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Proteinase K |
200 µL |
500 µL |
-20°C |
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Shipping: Dry Ice Shelf life: 2 years |
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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.