Animal Tissue Direct PCR Kit (for PCR series)

Product Information

Product Description/Introduction

The product is specially designed for amplification directly from animal tissues. Applicable samples include fresh or frozen mammalian viscera, nails, hair, zebrafish, drosophila, cultured cells, etc. The unique Animal Tissue Lysis Buffer A combine with Animal Tissue Lysis Buffer B to quickly release genomic DNA in micro tissues, without the need of genome extraction and purification, which is easy to operate, reduces the loss of samples, and greatly shortens the time-consuming experiments. The lysis product can be stored at -20 for a long time. 

2×Animal Tissue Direct PCR Mix with Dye contains all components required for PCR except templates and primers. The DNA Polymerase is capable of amplifying rapidly and resist inhibitors after modification. The PCR Mix also contains a variety of reaction enhancers, which can further improve the amplification efficiency and fidelity, also amplify target genes up to 3 kb.

Storage and Shipping Conditions

Ship with wet ice; Animal Tissue Lysis Buffer A, stored at 4; Other components shall be stored at -20; Valid for 12 months.

Product Contents

Assay Protocol / Procedures

1. Lysis of samples to release genomic DNA

a). Recommended sample dosage

b). Prepare Lysis Solution

a. Animal Tissue Lysis Buffer A appear white precipitates under low temperature. Incubate at 37 for 2-3 min until the white precipitate disappears before use.

c). Mix gently and centrifuge instantaneously 

d). Samples were weighed according to the recommended sample volume and placed in 1.5 mL EP tubes, to which the lysate required for individual samples was added, ensuring that the tissue was submerged in the lysate, and the tubes were centrifuged and incubated at 50°C for 10 min, followed by incubation at 95°C for 3 min.

(1) e). Centrifuge the lysed remains at 12000 rpm for 1 min to the bottom of the tube, and the supernatant can be used directly as a template for PCR reaction. The supernatant can be transferred to another sterile EP tube and placed at -20°C for long-term storage. 

(2) Recommended PCR reaction system (20 μL):

a：The dosage of template shall not exceed 1/10 of the reaction system.

b：Primer final concentration rang 0.5-1.0 μM. The recommended primer concentration is 0.5 μM. Too few primers may lead to low amplification yield or no amplification, and too many primers may lead to non-specific amplification.

(3) Recommended PCR amplification conditions:

Note

1. Before using the reagent, please gently mix it up and down.

2. Avoid freeze-thaw cycles, otherwise the genome will be degraded and the PCR amplification effect will be affected.

3. Animal Tissue Lysis Buffer A and Animal Tissue Lysis Buffer B should be used as they are prepared and left in place for a long period of time, which may affect lysis.

4. For samples that are difficult to lysis, such as nails, hair, shell, etc., the lysis time can be extended to 20 minutes at 50.

5. Before lysis of intestinal tissue, use ddH2O to clean the surface impurities, and the amplification effect is better.

6. After lysis, incomplete lysis tissue remains in EP tube, which is a normal phenomenon and does not affect the use.

For Research Use Only!

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