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Buccal Swab/FTA Card DNA Isolation Kit Genomic DNA Extraction or Purifiaction Kit from Buccal Swabs

Kit Description:

Quickly purify high-quality genomic DNA from buccal swab/FTA Card samples.

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

 Fast speed: Foregene Protease has higher activity than similar proteases, and digests tissue samples quickly; the operation is simple, and the genomic DNA extraction operation can be completed within 20-80 minutes.

◮ Convenient: The centrifugation is performed at room temperature, and there is no need for 4°C low-temperature centrifugation or ethanol precipitation of DNA.

◮ Safety: No organic reagent extraction is required.

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

◮ Micro-elution system: It can increase the concentration of genomic DNA, which is convenient for downstream detection or experiment.

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Product Detail

Product Tags

FAQ

Description

This kit provides an efficient and rapid method to obtain high-concentration genomic DNA from buccal swabs and FTA Card (blood spots). Using our companys unique DNA-only silica membrane spin column and formula, combined with Foregene Protease, high-concentration, high-quality genomic DNA can be extracted in 80 minutes. The specially designed small purification column binds genomic DNA, and the DNA can be eluted with a small amount (15μl) elution system to increase the concentration of the obtained genomic DNA, which is convenient for downstream detection or experiment. The kit can process one or more samples at a time, and the purification process does not require extraction of organic substances such as phenol, chloroform, and time-consuming isopropanol or ethanol precipitation, and the operation is simple and time-saving.

Specifications

50 Preps

Kit components

Buffer ST1

 Buffer ST2
 Linear Acrylamide
 Buffer PW
 Buffer WB
 Buffer EB
 Foregene Protease
 DNA-Only Column

Instructions

Features&advantages

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

-Fast speed: Foregene Protease has higher activity than similar proteases, digests samples quickly; simple operation.

-Convenient: The centrifugation is performed at room temperature, and there is no need for 4°C low-temperature centrifugation or ethanol precipitation of DNA.

-Safety: No organic reagent extraction is required.

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

-Micro-elution system: It can increase the concentration of genomic DNA, which is convenient for downstream detection or experiment.

Kit application

It is suitable for purification of genomic DNA from the following samples: buccal swabs, FTA Card (blood stains).

Storage and Shelf life

-This kit can be stored for 12 months under dry conditions at room temperature (15-25°C); if it needs to be stored for a longer period of time, it can be stored at 2-8°C.

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°C water bath for 10 minutes to dissolve the precipitate, and mix it before use.

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


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  • The purification column is clogged

    In this kit, in the genomic DNA extraction operation, the purification column is directly adsorbed on the sample enzymatic lysis mixture without centrifugation step, and the purification column may be blocked due to incomplete enzymization and high viscosity of the sample.

    The following possible causes are as follows:

    1. Incomplete enzymatic digestion of tissue samples.

    Recommendation: The sample processing time of Foregene Protease can be appropriately extended or the supernatant can be taken after centrifugation at 12,000 rpm (~13,400 × g) for 5 min.

    2. Excessive use of tissue samples or large tissues.

    Recommendation: It is best not to exceed 1 Buccal swab in the sample; if the sample is too large, increase the dosage of Buffer ST1, Foregene Protease, buffer ST2 accordingly.

    3. The sample viscosity is too high.

    Recommendation: Samples can be appropriately diluted with 10 mM of Tris-HCl before genomic DNA extraction.

    4. Fragments of the blood card have been sucked.

    Recommendation: The transient centrifugation time of step 6 of blood spot (FTA Card) genomic extraction can be appropriately extended.

    Low yield or no DNA

    There are often a variety of factors that affect genomic DNA yield, including sample origin, sample storage conditions, sample preparation, manipulation, etc.

    Genomic DNA cannot be obtained during extraction

    The possible causes are as follows:

    1. Improper preservation of samples or storage for too long leads to genomic DNA degradation.

    Recommendation: Oral swabs should preferably be freshly sampled, and it is not advisable to use preserved swabs for genomic DNA extraction operations; blood spot samples should ensure that the quality is qualified and the storage time should not be too long.

    2. Too little tissue usage may result in no extraction of the corresponding genomic DNA.

    Recommendation: Follow the buccal swab sampling instructions in the operation guide, and wipe as many times as possible so that enough cells can be attached to the oral swab for genomic DNA extraction; for blood spot sample extraction, the blood spot cutting area can be appropriately increased.

    3. Foregene Protease is improperly preserved, resulting in a decrease in its activity or inactivation.

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

    4. Improper preservation of the kit or storage time is too long, resulting in the failure of some components in the kit.

    Recommendation: Purchase a new Buccal swab DNA isolation kit for related procedures.

    5. Buffer WB does not add absolute ethanol.

    Recommendation: Confirm that buffer WB adds the correct volume of absolute ethanol.

    6. The eluent is not correctly added to the silicone film.

    Recommendation: Add 65 °C pre-warmed eluent drops to the middle of the silicone membrane and leave at room temperature for 5 min to increase the elution efficiency.

    Low-yield genomic DNA isolated

    The following possible causes are as follows:

    1. Improper preservation of samples or storage for too long leads to genomic DNA degradation.

    Recommendation: Oral swabs are preferably freshly sampled, and preserved swabs should not be used for genomic DNA extraction.

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

    Recommendation: Follow the oral swab sampling instructions in the operating guide, wiping as many times as possible so that enough cells can be attached to the oral swab for genomic DNA extraction.

    3. Foregene Protease is improperly preserved, resulting in a decrease in its activity or inactivation.

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

    4. Eluent problems.

    Recommendation: Use Buffer EB for elution; if using ddH2O or other eluents, confirm that the pH of the eluate is between 7.0-8.5.

    5. The eluate is not correctly added dropwise.

    Recommendation: Add eluent drops to the middle of the silicone membrane and leave at room temperature for 5 min to increase elution efficiency.

    6. The elution liquid accumulates too little.

    Recommendation: Use eluent for genomic DNA elution as required in the instructions, at least no less than 15 μl.

    Low purity of genomic DNA isolated

    Low genomic DNA purity can lead to failure or unsatisfactory results of downstream experiments, such as: enzymes cannot be cut open, PCR can not get the gene fragment of interest, etc.

    The possible causes are as follows:

    1. Heteroprotein pollution, RNA pollution.

    Analysis: The purification column was not washed using Buffer PW; the Buffer PW wash purification column was not washed using the correct centrifugal speed.

    Recommendation: Ensure that there is no precipitation in the supernatant before adding ethanol; be sure to wash the purification column according to the instructions, and this step cannot be omitted.

    2. Impurity ion pollution.

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

    Recommendation: Be sure to wash Buffer WB 2 times as directed to remove residual ions as much as possible.

    3. RNA enzyme contamination.

    Analysis: Foreign RNases were added to buffer; Buffer PW wash operation was incorrect, resulting in RNase residues, affecting downstream RNA experimental operations, such as in vitro transcription.

    Recommendation: Foregene series nucleic acid isolation kits can remove RNA without additional addition of RNase,thus buccal Swab/FTA Card DNA Isolation Kit needn’t add RNase; be sure to follow the instructions for Buffer PW washing purification column, and this step cannot be omitted.

    4. Ethanol residue.

    Analysis: Buffer WB did not perform empty tube centrifugation after washing the purification column.

    Recommendation: Perform the correct empty tube centrifugation operation according to the instructions.

    5. Other impurity pollution.

    Analysis: Saved samples or special samples are not pretreated.

    Recommendation: Thoroughly pretreat the sample as instructed.

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