Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • ATAQ-Seq qPCR troubleshooting

    Hello,

    I am attempting to perform ATAQ-seq on human macrophages. I have performed the protocol, though have encountered an issue at the qPCR step (after the initial 5 cycle amplification step). I find i have no amplification at all.

    I was worried it was because i lost my cell pellet during the initial wash stages so i have nanodropped my samples at various steps:

    1. After the initial purification following the transposition reaction
    2. After the initial 5 cycle PCR amplification step
    3. After final purification step

    However, i have DNA at all of these steps, indicating i haven't lost my pellet and the DNA is being amplified.

    For this protocol i am using the Illumina Nextera Index Kit (FC-121-1012) as the primers.
    Regarding this i have two questions:
    1. Are these indices compatible with qPCR?
    2. For each sample should use the same index pair used in the amplification PCR in the qPCR?

    Any other suggestions would be greatly appreciated!
    Thanks

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
8 views
0 likes
Last Post seqadmin  
Started by seqadmin, Yesterday, 06:07 PM
0 responses
8 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