Animal Tissue DNA Isolation Kit DE-05011/05012/05013 For Genomic DNA Purification

Animal Tissue DNA Isolation Kit Cat.No.DE-05011/05012/05013 For Genomic DNA Purification From Cultured Cells And Animal

Animal Tissue DNA Isolation Kit

Cat.No.DE-05011/05012/05013

For genomic DNA purification from cultured cells and animal tissues

Product Descprition:

This kit uses a DNA-only column that can specifically bind DNA, Foregene protease and a unique buffer system. High-quality genomic DNA can be extracted from various cultured cells and animal tissues within 30 to 50 minutes.

The DNA-only silica gel membrane used in the spin column is Foregene's unique new material, which can effectively and specifically bind to DNA, and maximize the removal of RNA, impurity proteins, ions and other organic compounds in cells. 5-80μg high-quality genomic DNA can be purified from 10-50mg tissue.

Product components:

Features&advantages:

-No RNase contamination: The DNA-Only Column provided in the kit makes it possible to remove RNA from genomic DNA without additional RNase during the experiment, thereby preventing the laboratory from being contaminated by exogenous RNase.

-Fast speed: Foregene Protease has higher activity than similar proteases and digests tissue samples faster;

-Simple: the genomic DNA purification operation can be completed in 50 minutes.

-Convenient: The centrifugation is performed at room temperature, 4℃ centrifugation and ethanol precipitation of DNA is not required.

-Safety: no organic reagent is used.

-High quality: The purified genomic DNA has large fragments, no RNA, no RNase, and extremely low ion content, which can meet the requirements of various experiments.

Applications of genomic DNA:

The genomic DNA purified by Animal Tissue DNA Isolation Kit has high purity and can be used for routine molecular biology operations, such as: enzyme digestion, PCR, Southern hybridization, library construction and other experiments.

Product Quality Control:

In accordance with FOREGENE's Total Quality Management System, each batch of animal tissue DNA isolation kits are strictly tested multiple times to ensure the reliability and stability of the quality of each batch of kits.

Genomic DNA extraction yield and purity:

The extraction yield of animal tissue genomic DNA is related to the tissue source, storage conditions, storage time, dosage and other factors. The purity of the obtained genomic DNA meets the routine molecular biology experimental operation, and its OD260/280 is between 1.7-1.9. The yield of genomic DNA extracted using this kit is shown in the following table:

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

- Sample should avoid repeated freezing and thawing, otherwise the extracted DNA fragments will be smaller and the extraction yield will be reduced.

- Before using the kit, carefully check whether there is precipitation in Buffer L1, Buffer L2 and Buffer PW. If there is precipitation, please dissolve it at 37℃, mix well before use.

- Before using the kit, be sure to check whether Buffer WB is added with anhydrous ethanol according to the instructions. Buffer WB was added with 60ml anhydrous ethanol (DE-05011), 120ml anhydrous ethanol (DE-05012) and 300ml anhydrous ethanol (DE-053013) before use.

- Elution volume: Buffer EB should not be less than 100μl, otherwise it will affect the DNA yield.

- Remember not to add RNase to any Buffer.

- All centrifugation steps are centrifuged at room temperature (15-25℃) using a desktop centrifuge.

- All experimental steps were carried out at room temperature (15-25℃).

Storage and Shelf life:

- The kit can be stored for 12 months at room temperature (15-25℃), or 2-8℃ for longer time.

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

- Foregene Protease solution has a unique formula and is active for a long time (3 months) at room temperature; its activity and stability will be better when stored at 4℃, so it is recommended to store it at 4℃, remember not to store it at -20℃ save.

DNA-Only Column characters

*:The minimum elution system of 100μl is a reasonable recommended volume given in consideration of DNA recovery rate and concentration. If in order to increase the yield of DNA, the volume of the eluate can be appropriately increased; if in order to increase the concentration of purified DNA, the volume of the eluate can be appropriately reduced on the premise of sacrificing a part of the DNA yield, such as using a 50μl elution system, in order to obtain higher concentrations of DNA.

Genomic DNA extraction yield and purity

The extraction yield of animal tissue genomic DNA is related to the tissue source, storage conditions, storage time, dosage and other factors. The purity of the obtained genomic DNA meets the routine molecular biology experimental operation, and its OD260/280 is between 1.7-1.9. The yield of genomic DNA extracted using this kit is shown in the following table:

Note: The data in this table is for reference only. In actual operation, the data obtained will be slightly different from the data in this table due to factors such as storage conditions and operating proficiency of the materials used.

Workflow

Problem Analysis Guide

The following is an analysis of the problems that may be encountered in the extraction of genomic DNA from tissue samples, hoping to be helpful to your experiments. In addition, for other experimental or technical problems other than operation instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us: 028-83360257 or E-mali: Tech@foregene.com.

Low yield or no DNA

There are usually multiple factors that affect the yield of genomic DNA, including sample source, sample storage conditions, sample pretreatment, manipulation, etc.

Genomic DNA could not be obtained during extraction

1. The tissue samples are improperly stored or stored for too long, resulting in the degradation of the genomic DNA.

Recommendation: Store tissue samples in liquid nitrogen or -80°C; try to use newly collected tissue samples for genomic DNA extraction.

2. Too little tissue consumption may lead to the failure to extract the corresponding genomic DNA.

Suggestion: For tissue samples that have been stored for a long time or have severe genomic DNA degradation, the amount of tissue samples can be appropriately increased in order to extract considerable genomic DNA. The amount of sample can be determined according to DNA needs, but it should not exceed 50mg.

3. Improper storage of Foregene Protease results in reduced or inactivated activity.

Recommendation: Confirm the storage conditions of Foregene Protease or replace it with a new Foregene Protease for enzymatic hydrolysis.

4. The kit is improperly stored or stored for too long, causing some components in the kit to fail.

Recommendation: Purchase a new animal tissue genomic DNA extraction kit for related operations.

5. Improper use of the kit. For example, some tissues need special kits for processing.

Recommendation: Purchasing a kit for samples, such as the extraction of soil genomic DNA, requires purchasing a dedicated Soil DNA Isolation Kit.

6. Buffer WB without adding anhydrous ethanol.

Recommendation: Make sure to add the correct volume of anhydrous ethanol to Buffer WB.

7. The eluent was not dripped onto the silica membrane properly.

Suggestion: Add the pre-heated eluent at 65°C dropwise to the middle of the silica gel membrane, and leave it at room temperature for 5 minutes to increase the elution efficiency.

Extracted genomic DNA with low yield

1. The sample is improperly stored or stored for too long, resulting in the degradation of genomic DNA.

Recommendation: Store tissue samples in liquid nitrogen or -80; try to use newly collected tissue samples for genomic DNA extraction.

2. If the amount of tissue samples is too small, the extracted genomic DNA will be less.

Suggestion: For tissue samples that have been stored for a long time or have severe genomic DNA degradation, the amount of tissue samples can be appropriately increased to obtain considerable genomic DNA. The amount of sample can be determined according to DNA needs, but it should not exceed 50mg.

3. Excessive amount of sample will lead to incomplete digestion and incomplete centrifugation. The purification column may be blocked during centrifugation on the column, and the extracted genomic DNA will be less.

Suggestion: According to different tissues, the dosage should be adjusted within 10-50 mg. When the digestion is incomplete or the purification column is blocked, please reduce the corresponding tissue dosage.

4. The sample is not preprocessed or preprocessed incompletely.

Suggestion: Specially preserved samples or some relatively special samples must be pretreated before enzymatic hydrolysis, which can remove the sample preservation solution or factors affecting protease activity, so that the enzymatic hydrolysis reaction can be carried out more thoroughly, and higher yield genomic DNA can be obtained.

5. Special preserved tissue samples, such as paraffin-embedded, rat tail, etc.

Recommendation: Purchase a dedicated genomic DNA extraction kit, such as paraffin-embedded samples, you can choose FFPE DNA Isolation Kit; mouse tail samples, you can choose Mouse Tail DNA Mini Kit. These tissues are treated with their special kits, and the DNA extraction yield will be higher and the effect will be more ideal.

6. Improper storage of Foregene Protease results in reduced or inactivated activity.

Recommendation: Confirm the storage conditions of Foregene Protease or replace it with a new Foregene Protease for enzymatic hydrolysis.

7. The eluent problem.

Recommendation: Please use Buffer EB for elution; if using ddH₂O or other eluents, make sure the pH of the eluent is between 7.0-8.5.

8. The eluent was not added dropwise correctly.

Suggestion: Please add the elution drop to the middle of the silica membrane and leave it at room temperature for 5 minutes to increase the elution efficiency.

9. The eluent volume is too low.

Suggestion: Please use the eluent for genomic DNA elution according to the instructions, at least not less than 100μl.

Extracted genomic DNA with low purity

The low purity of genomic DNA will lead to the failure or unsatisfactory effect of downstream experiments, such as: the enzyme cannot be cut, and the target gene fragment cannot be obtained by PCR.

1. Miscellaneous protein contamination, RNA contamination.

Analysis: Buffer PW was not used to wash the column; Buffer PW was not used to wash the column at the correct centrifugation speed.

Recommendation: Try to ensure that there is no precipitation in the supernatant when the supernatant is passed through the column; be sure to wash the purification column with Buffer PW according to the instructions, and this step cannot be omitted.

2. Impurity ion pollution.

Analysis: The Buffer WB wash column was omitted or only washed once, resulting in residual ionic contamination.

Recommendation: Be sure to wash twice with Buffer WB according to the instructions to remove residual ions as much as possible.

3. RNase contamination.

Analysis: Exogenous RNase is added to the Buffer; incorrect Buffer PW washing operation results in residual RNase, which affects downstream RNA experimental operations, such as in vitro transcription.

Suggestion: Foregene series nucleic acid extraction kits can remove RNA without additional RNase, and all reagents in Animal Tissue DNA Isolation Kit do not need RNase; be sure to wash the purification column with Buffer PW according to the instructions, and this step cannot be omitted.

4. Ethanol residues.

Analysis: After washing the purification column with Buffer WB, no empty tube centrifugation was performed.

Recommendation: Follow the instructions for proper empty tube centrifugation.

5. Other impurities pollution.

Analysis: Saved samples or special samples were not preprocessed.

Recommendation: Thoroughly pre-treat the sample according to the operating instructions.