Catalog No: K0568600
A number of applications:
- 1 set
- 40 reactions at 50 ul PCR volume / 100 reactions at 20 ul PCR volume
- 1 year at -20°C, 1 month at 4°C
- (Product may be shipped on blue ice and should be stored immediately at -20°C.)
The EzWay Mouse Tail Direct PCR Kit allows the amplification of DNA directly from a mouse tail without DNA extraction. Simply cut the tail of the mouse and incubate it in the lysis buffer provided in the kit for 10 min and then use 1 ul of lysate for PCR. it doesn’t require overnight incubation or proteinase K (PK) treatment.
- Fast and simple method for genotyping transgenic mice without DNA purification
- Only 10 min incubation is required.
- It is not necessary to add Proteinase K
- Saving enormous amounts of material and time.
- Optimized MasterMix type containing EzWayTM Hot Taq PCR enzyme, dNTP, forward PCR buffer, MgCl2, red dye, and additives
- Direct loading of PCR products without adding red dye
1. Cut the tail of the mouse to a length of 1-2 mm (or 5-10 mg) and add lysis buffer. Incubate the tube at 60°C for 10 min in a water bath/rotary hybridization oven or PCR thermal cycler (Vortex will sometimes be useful for lysis). DNA molecules from the tail tissue are released.
2. Vortex and spin down briefly.
3. Directly use 1 ul of crude lysate supernatant for every 50 ul of PCR reaction.
Note: Mouse Tail Direct Lysis Buffer contains a high concentration of EDTA and detergents, therefore, higher amounts of lysate may affect the buffer’s ability to neutralize inhibitors.
- ezWay Direct Hot Taq PCR MasterMix contains 3.0 mM MgCl2 (final 1.5 mM MgCl2). Generally, 1.5 mM MgCl2 can give satisfactory PCR results, but for MgCl2 greater than 1.5 mM, add MgCl2 separately.
- Magic Buffer will improve the amplification of DNA from templates that have high G+C content and a high degree of secondary structure. We recommend that the added volume should not exceed 25% (v/v) of the final PCR volume.
- Mix gently.
- When using a thermal cycler without a heated lid, add approximately 100 ul of
mineral oil on top of the mixture.
- Perform thermal cycling.
a. Primers should be 15 to 30 bases long and have a G+C content close to 50%.
b. Magic Buffer will improve the amplification of DNA from templates that have a high G+C content and a high degree of secondary structure. We recommend that the volume added should not exceed 25% (v/v) of the final PCR volume.
c. If chemically modified hot start Taq PCR enzyme is used, the best initial.denaturation time is 10-15 minutes at 95°C.
- Amplified DNA can be detected by various electrophoresis techniques. The most common techniques are agarose or polyacrylamide gel electrophoresis according to the size of the amplicon. TAE gel is highly recommended because the bands can be detected as scattered and/or distorted in TBE gel.