Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • MiSeq sequencing primer question

    We are trying to do single-cell TCR sequencing using Mark Davis protocol based on his 2015 Nature Biotech publication. The link for the paper is:

    We used the same primer set as the paper. The PE primers for the final PCR reaction are:
    PEprimer1: AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT
    PEprimer2: AAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT

    My questions are:
    1. I can see PEprimer1 contains P5 sequence primer. What is PEprimer2? What should I use for the other end of sequence primer?
    2. For the final (third) PCR reaction, do we have to use PAGE purified PEprimer1 and PEprimer2? Do we need to order PE primers modified with phosphorothioate bonds between the two last (3') nucleotides according to Illumina’s Nature Method paper? Or this is just for the sequencing part?

    Thank you so much!

  • #2
    Hi enum,
    Did you succeed with single-cell TCR sequencing?
    I just ordered Mark Davis primes. I tried Smarter kit with not much success.
    I'll appreciate your help.

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Best Practices for Single-Cell Sequencing Analysis
      by seqadmin



      While isolating and preparing single cells for sequencing was historically the bottleneck, recent technological advancements have shifted the challenge to data analysis. This highlights the rapidly evolving nature of single-cell sequencing. The inherent complexity of single-cell analysis has intensified with the surge in data volume and the incorporation of diverse and more complex datasets. This article explores the challenges in analysis, examines common pitfalls, offers...
      Yesterday, 07:15 AM
    • seqadmin
      Latest Developments in Precision Medicine
      by seqadmin



      Technological advances have led to drastic improvements in the field of precision medicine, enabling more personalized approaches to treatment. This article explores four leading groups that are overcoming many of the challenges of genomic profiling and precision medicine through their innovative platforms and technologies.

      Somatic Genomics
      “We have such a tremendous amount of genetic diversity that exists within each of us, and not just between us as individuals,”...
      05-24-2024, 01:16 PM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Yesterday, 08:18 AM
    0 responses
    13 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, Yesterday, 08:04 AM
    0 responses
    12 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 06-03-2024, 06:55 AM
    0 responses
    13 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 05-30-2024, 03:16 PM
    0 responses
    27 views
    0 likes
    Last Post seqadmin  
    Working...
    X