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Instruction Manual:

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Personlized Products China Bacteria Genomic DNA Extraction and Viral Rna Extraction Kit, We have many years’ experience in hair product production, and our strict QC Team and skilled workers will ensure that we provide you with top hair solutions with the best hair quality and workmanship. You will get successful business if you choose to cooperate with such a expert manufacturer. Welcome your order cooperation!

Problem Analysis Guide

The following is an analysis of the problems that may be encountered in the extraction of plant genomic DNA, 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 many factors that affect the yield of genomic DNA, including the source of the sample, the age of the sample, the storage conditions of the sample, and the operation.

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 -20°C; try to use newly collected samples for genomic DNA extraction.

2. Too little sample amount may cause the corresponding genomic DNA not to be extracted.

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 the sample can be determined according to the DNA needs, but the fresh sample should not exceed 100mg, and the dry sample should not exceed 30mg.

3. The sample is not ground with liquid nitrogen or placed for too long after liquid nitrogen.

Suggestion: During DNA extraction, the sample needs to be fully ground with liquid nitrogen to break the cell wall; after grinding, please transfer the sample powder to PL1 preheated at 65°C as soon as possible (once the ground powder is melted, the genomic DNA will begin to degrade rapidly) .

4. 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.

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

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

6. Improper use of the kit.

Suggestion: Purchase a Plant DNA Isolation Kit dedicated to samples for extraction and purification of plant genomic DNA.

7. Buffer WB without adding anhydrous ethanol.

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

8. The eluent was not dripped onto the silica membrane correctly.

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

Extraction to obtain low-yield genomic DNA

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

Recommendation: Store tissue samples at -20℃; 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: Some plant samples are rich in water, such as aquatic plants such as algae, etc., the dosage can be appropriately increased or the water can be dehydrated a little before the operation.

3. The samples were not thoroughly ground with liquid nitrogen or were left at room temperature for too long after grinding.

Suggestion: The liquid nitrogen grinding must be sufficient, and the sample cell wall should be broken as much as possible; immediately after the grinding, the sample powder should be transferred to 65℃ preheated Buffer PL1 for the next step.

4. Not using the correct kit.

Recommendation: Use a dedicated Plant DNA Isolation Kit to extract and purify plant genomic DNA.

5. 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.

6. 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.

7. The eluent is not dripped 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.

8. The eluent volume is too small

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 poor 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.

Suggestion: 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 washing operation in Buffer PW will result in residual RNase and affect 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 Plant 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.

Instruction Manual:

Plant DNA Isolation Kit Instruction Manual