Lab Reagent Animal Tissue Direct PCR Kits Performing PCR Directly From Animal Tissue

Lab reagent Animal Tissue Direct PCR Kits performing PCR directly from animal tissue without prior DNA purification

Product Descprition

The Animal Tissue Direct PCR kit uses a unique lysis buffer system to rapidly release genomic DNA from animal tissue samples for PCR reactions, so it is especially suitable for large-scale genetic testing.

The process of releasing genomic DNA from the lysis buffer is completed within 10-30 minutes at 65°C, and the released trace amount of DNA can be used as a template for PCR reaction without other processes such as protein removal and RNA removal.

2×PCR Easy^TM Mix has a strong resistance to PCR reaction inhibitors, and can use the lysate of the sample to be tested as a template for efficient and specific amplification. This reagent contains Foregene D-Taq DNAPolymerase, dNTPs, MgCl₂, reaction buffer, PCR optimizer, and stabilizer. It can be used in combination with lysis buffer to detect samples quickly and easily, and has the characteristics of high sensitivity, strong specificity and good stability.

D-Taq DNA polymerase is a DNA polymerase specially developed by Foregene for direct PCR reactions.

D-Taq DNA polymerase has strong tolerance to a variety of PCR reaction inhibitors, and can efficiently amplify trace amounts of DNA in various complex reaction systems. The amplification speed can reach 2Kb/min, which is especially suitable for Direct PCR reaction.

Product components:

Features&advantages:

- No need for time-consuming and expensive DNA purification.

- The sample requirement is small, as little as 5 mg of animal tissue can be used for experiments.

- No special treatment such as grinding and crushing is required, and the operation is simple.

- The optimized PCR system makes PCR more specific and more resistant to PCR reaction inhibitors.

Application:

- Scope of application: a variety of animal tissues.

- DNA released from sample lysis: Use as PCR template only.

- The kit can be used for the following purposes: identification of transgenes, animal genotyping, etc.

Product quality control:

According to FOREGENE's Total Quality Management System, each batch of animal direct PCR kits is strictly tested multiple times to ensure the reliability and stability of the quality of each batch of kits.

Kit Component Information

Buffer AL: Provides the environment required for animal tissue lysis reactions.

Foregene Protease: Lyses animal tissue in the presence of lysis buffer to release genomic DNA.

2×PCR EasyTM Mix: Contains D-Taq DNA Polymerase specially modified by Fuji Bio, dNTPs, MgCl2, reaction buffer, PCR reaction enhancer, optimizer and stabilizer, etc. For PCR reactions, simply add the appropriate lysis mix, primers, and ddH2O to the 2×PCR EasyTM Mix for PCR reactions.

6×DNA Loading Buffer: This Loading Buffer does not contain SDS. It is recommended to use the 6× DNA Loading Buffer provided with the kit when performing agarose gel electrophoresis in order to obtain good electrophoresis results. Do not use Loading Buffer containing SDS, otherwise there will be a large tail of light in the lane during electrophoresis, which will affect the experimental results.

Storage conditions

1. Shipping Conditions

The whole process is transported in a low temperature ice box to ensure that the Part I and Part II of the kit are in a state of <4°C.

2. Storage conditions

-This kit Part I is stored at room temperature or 2-8℃.

-Reagent Buffer AL can be stored for 12 months under dry conditions; for longer storage, it can be stored at 2–8°C.

Note: If stored at low temperature, the solution is prone to precipitation. Be sure to place the solution in the kit at room temperature for a period of time before use, and if necessary, preheat in a 37°C water bath for 10 minutes to dissolve the precipitate, and mix well before use.

-Reagent Foregene Protease has a unique formula, in order to ensure its better activity and stability, please store it at 4℃.

-Reagent 6×DNA Loading Buffer, can be stored at 4°C or -20°C for a long time.

This kit Part II is stored at -20°C.

-Reagent 2×PCR Easy^TM Mix, if used frequently, it can also be stored at 4°C for a short time (limited to be used up within 10 days).

Precautions: (Please be sure to read the precautions carefully before using the kit)

- Pay attention to the cleanliness of laboratory equipment and the operation method of the experiment to avoid cross-contamination between samples.

- Please try to use freshly collected animal tissue samples for experiments. If the tissue samples are stored for a long time, please avoid repeated freezing and thawing of the samples.

- If there is precipitation in Buffer AL, it can be placed at 37°C until the precipitation disappears, and the solution is shaken before use.

- Foregene Protease has a unique formula, please store at 4℃, never -20℃.

- 2×PCR Easy^TM Mix should avoid repeated freezing and thawing, otherwise it will affect the PCR efficiency.

- If the ambient temperature is too high, the 2×PCR EasyTM Mix may become turbid. It can be placed on ice for 1-2 minutes. When the solution is clear, invert it up and down for 3-5 times before use.

- Do not use Loading Buffer containing SDS during electrophoresis detection, otherwise a large group of trailing bright bands will appear in the swimming lane during electrophoresis, which will affect the experimental results.

PCR control reaction

In the analysis of PCR results, whether positive or negative, without a control reaction, we cannot be sure that the results are reliable. In order to facilitate the analysis of subsequent experimental results, we recommend setting positive and negative PCR control reactions during PCR in order to exclude the interference of false positives or false negatives.

A: Positive control

Use primers of conserved genes in the sample that are easy to amplify and use purified sample DNA as a template to perform a positive control reaction to determine the correctness of the PCR reaction system and conditions and the effectiveness of 2×PCR Easy_TM Mix. The preparation of the reaction system is shown in Table 3 below.

Table 3:Control PCR reaction system preparation

1*: Primers of conserved genes that are easier to amplify in this sample can be selected, such as β-Actin gene, conserved sequences in the genome, etc. (please ensure the availability of these primers).

2*: The DNA purified from the sample can be selected, or it can be selected according to the experimental needs.