Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Primers for ZCCHC24

    Dear Seqanswer members,

    I am desperately trying to amplify/sequence the first exon of the ZCCHC24 gene. I tried it with the following primers

    Fwd 5' cgaggcagatagcggatcg 3'

    Rev 5' cttggcttggcttcgttcaac 3'

    with several polymerases (PeqLab Taq-DNA-Polymerase, TaKaRa rTaq, TaqOne GC Kit) at several annealing temperatures (ranging from 50°C to 60°C). I also tried to add 1 µl DMSO. I get either unspecific products (which do not contain a product of desired length) or there is no product at all.

    A former employee already have tried the following primers and failed as well:

    Fwd 5' aactttcgctgcctccttc 3'

    Rev 5' ctgtctgacccgctcttcct 3'

    I don't know which polymerases and programms she used though.

    Do you have any ideas, how I could amplify/sequence this exon? Any suggestions, including primer suggestions are appreciated!

    Thank you in advance,

    Anna.

  • #2
    Hi, Anna. What is your target species?

    Comment


    • #3
      Hi greekkey,

      I solved the problem in the meantime by adding DMSO, BSA and Betaine.

      Thank you for trying to help me though!

      Best,
      Anna.

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Current Approaches to Protein Sequencing
        by seqadmin


        Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
        04-04-2024, 04:25 PM
      • 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

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, 04-11-2024, 12:08 PM
      0 responses
      25 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      27 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      24 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-04-2024, 09:00 AM
      0 responses
      52 views
      0 likes
      Last Post seqadmin  
      Working...
      X