Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • DNA extraction from Saliva

    Hi everybody!!
    I have to extract DNA from Saliva, in order to realize some ION Torrent PGM libraries. I'd like to ask if you know which is the best kit for DNA extraction. In the past we used the Oragene extraction protocol, but our samples were full of proteins. So we have thought about using the DNA mini blood kit, that is compatible also with saliva. In fact our aim is to have a good concentration, but also a great quality of DNA samples. One of the limits of this kit is the cost, because it's really expensive. So I tried to find out something similar, but less expensive. I've read something about the NORGEN DNA saliva isolation kit. Have you ever used it? Do you know other kits that I can use?
    Thank you in advance for answering!
    Anna

  • #2
    My lab always uses the Qiagen DNeasy blood & tissue kit for extracting DNA from tissues. We used it for prepping DNA for Illumina sequencing and it worked well.

    Comment


    • #3
      We've made NGS libraries from Oragene DNA and 23andMe used it for my genome. Oragene is expensive but you get masses of high-quality DNA.

      here's a list of providers I am aware of:
      Oragene. http://www.dnagenotek.com/US/product...ctionkits.html
      Norgen. http://norgenbiotek.com/display-product.php?ID=493
      nexttec 1-Step. http://www.nexttec.biz/products
      PSP SalivaGene http://www.invitek.de/products_and_s...agene_dna_kit/

      I'd be intersted to hear if anyone has compared saliva extractions kits.

      Comment


      • #4
        Hi Anna

        We've used Oragene for NGS on both the Ion Torrent and Illumina systems (GAII and HiSeq2000), both targeted and whole genome and we didn't have any problems. We always extract using prepIT-L2P (formerly Oragene Purifier), an EtOH precipitation-based method, and we don't have problems with protein contamination.

        Are you assessing protein contamination by 260/280 ratio? If so, you should be able to obtain 1.7 or higher in most cases with a prepIT-L2P extraction. Column-based extraction like prepIT-C2D or various Qiagen kits might give you a bit higher ratio (1.75 or 1.8) but you'll sacrifice yield and, in my experience, it's not necessary to use column extractions for NGS.

        Mike Tayeb
        Manager, Technical Support
        DNA Genotek (manufacturers of Oragene)

        Comment


        • #5
          We've substituted saliva into Qiagen whole blood preps with good results, but be aware of how much bacterial DNA will be in saliva preps.

          Comment


          • #6
            Saliva Devices

            Hi guys,

            Often times the collection can be the key to getting a pure and high quality sample. I work for a company that produces a range of saliva collection devices, I think you might find the DNA·SAL™ : http://4saliva.com/products/dna-sal/ a perfect fit for gathering a good, clean, and simple sample.

            We are a small company really focused on customer service, we are really happy to help anyone through any issues or questions they might have. We go to great lengths to customize our products where there is need as well. Hope this helps.

            Comment


            • #7
              There's a NEB kit that removes human DNA for microbiome workflows; NEB has a protocol to elute the human DNA.

              Facilitates the enrichment or exclusion of microbial DNA from samples containing methylated host DNA (including human).


              Comment

              Latest Articles

              Collapse

              • seqadmin
                Strategies for Sequencing Challenging Samples
                by seqadmin


                Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
                03-22-2024, 06:39 AM
              • seqadmin
                Techniques and Challenges in Conservation Genomics
                by seqadmin



                The field of conservation genomics centers on applying genomics technologies in support of conservation efforts and the preservation of biodiversity. This article features interviews with two researchers who showcase their innovative work and highlight the current state and future of conservation genomics.

                Avian Conservation
                Matthew DeSaix, a recent doctoral graduate from Kristen Ruegg’s lab at The University of Colorado, shared that most of his research...
                03-08-2024, 10:41 AM

              ad_right_rmr

              Collapse

              News

              Collapse

              Topics Statistics Last Post
              Started by seqadmin, Yesterday, 06:37 PM
              0 responses
              10 views
              0 likes
              Last Post seqadmin  
              Started by seqadmin, Yesterday, 06:07 PM
              0 responses
              9 views
              0 likes
              Last Post seqadmin  
              Started by seqadmin, 03-22-2024, 10:03 AM
              0 responses
              49 views
              0 likes
              Last Post seqadmin  
              Started by seqadmin, 03-21-2024, 07:32 AM
              0 responses
              67 views
              0 likes
              Last Post seqadmin  
              Working...
              X