Technical

  1.  
  2. What is the capacity of the binding columns?
    The capacity of the Fast ID binding columns is approximately 60 microgram DNA but the typical yield based on 200 mg starting material is 10 microgram DNA.
     
  3. What is the expected recovery of genomic DNA using the Fast ID kit?
    Expected recovery should be ~ 90%.  If it is critical to recover the maximum yield of DNA, then a second 1xTE elution step can be added to the procedure to recover any remaining DNA still bound to the column.
     
  4. What are the typical DNA yields obtained with the Fast ID kit from a variety of plant, animal, and bacterial samples?
    See the Table included in the Product Data section.
     
  5. Is the Fast ID kit able to successfully isolate DNA from processed foods and/or animal hides such as leather?
    Yes. The Fast ID Genomic DNA extraction kit can purify DNA from most samples, provided the DNA is not too fragmented.  Leather has an additional complication in that it is processed with strong chemicals which may result in PCR inhibition.
     
  6. Is the Fast ID Genomic DNA Extraction kit able to extract DNA from specimens that were originally fixed in formalin and then stored in 70% ethanol?
    The Fast ID kit is able to isolate and purify DNA from such specimens.
     
  7. Is the Fast ID kit able to isolate DNA from Gram positive bacteria?
    Yes, but lysozyme enzyme should be added to the lysis buffer. Lysozyme is not provided with the Fast ID kit and must be purchased separately.  

Procedural


  1. The DNA yield was not what I expected.  How can I increase DNA yield using the Fast ID kit?
    A second 1xTE elution step can be added to the procedure to recover any remaining DNA still bound to the column.  In addition, increasing the amount of Proteinase K added during the lysis step may increase the yield of DNA from certain protein-rich samples. Finally, sample size can be as large as 1g as long as the volume of lysis buffer is increased correspondingly in the lysis step.
       
  2. What kind of centrifuge do you recommend using with the Fast ID 96-well kit?
    A centrifuge (and rotor) capable of accommodating 96-well deep plate carriers is necessary.  
     
  3. Are there any necessary pretreatments for paraffin embedded tissues?
    Yes.  Tissues embedded in paraffin should first be treated with xylene to remove paraffin prior to DNA extraction using the Fast ID kit.
     
  4. I often observe PCR inhibition with DNA extracted using the Fast ID kit.  How can I avoid this?
    PCR inhibition can be due to salt carry-over in the DNA from the buffer. It is very important to ensure proper washing with 75% ethanol during the ethanol washing steps. Always fill the DNA binding column to the rim with 75% ethanol to ensure removal of any salt. PCR inhibition can also occur because the sample type itself is rich in inhibitory compounds. In this case, an additional washing step with Genomic Wash buffer is recommended. Alternatively, a chloroform purification step can be helpful (see Troubleshooting section of protocol).  

Applications

  1.  
  2. Is the Fast ID kit suitable for forensic applications? 
    We do not recommend that DNA obtained using the Fast ID kit be used for forensic analysis.  There are a number of commercially available kits specifically designed for that purpose.