Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • gilly
    replied
    Hi.

    Yes we do see them occasionally I believe the cause is amplification primer dimmers. If you take these libraries into emPCR then you run the risk of getting high enrichment with an increase in very short or even null beads.

    This peak can be removed by performing a 4-cycle PCR, using identical conditions to those listed in the paired end manual (except with the cycle number reduced to 4 obviously). This step is to double-strand all of your products. This is then followed by a standard SPRI clean up (1.8x SPRI to sample ratio, as per Agencourt’s protocol).

    Leave a comment:


  • 454User
    started a topic LMW band in paired end libraries?

    LMW band in paired end libraries?

    Hello all,

    To those of you that prepare Ti 3 and 8 kb PE libraries, have you ever experienced a prominent, low molecular weight band around 100-110 nt in your ssDNA product? Please see the attached images for examples.

    I have seen this band without fail in many PE preps that span reagent lots and operator experience. While its abundance relative to the major library band varies, its presence does not. I am of the mind that this band represents:

    1.) Library adaptor dimers (though these adaptors are not phosphorylated and I would not expect PNK to carry through the end polish cleanup)

    or

    2.) Amplification primer dimers that display this size due to their %GC

    Any comments or insight would be greatly appreciated!

    -J
    Attached Files

Latest Articles

Collapse

  • seqadmin
    Exploring the Dynamics of the Tumor Microenvironment
    by seqadmin




    The complexity of cancer is clearly demonstrated in the diverse ecosystem of the tumor microenvironment (TME). The TME is made up of numerous cell types and its development begins with the changes that happen during oncogenesis. “Genomic mutations, copy number changes, epigenetic alterations, and alternative gene expression occur to varying degrees within the affected tumor cells,” explained Andrea O’Hara, Ph.D., Strategic Technical Specialist at Azenta. “As...
    07-08-2024, 03:19 PM
  • seqadmin
    Exploring Human Diversity Through Large-Scale Omics
    by seqadmin


    In 2003, researchers from the Human Genome Project (HGP) announced the most comprehensive genome to date1. Although the genome wasn’t fully completed until nearly 20 years later2, numerous large-scale projects, such as the International HapMap Project and 1000 Genomes Project, continued the HGP's work, capturing extensive variation and genomic diversity within humans. Recently, newer initiatives have significantly increased in scale and expanded beyond genomics, offering a more detailed...
    06-25-2024, 06:43 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by seqadmin, 07-16-2024, 05:49 AM
0 responses
30 views
0 likes
Last Post seqadmin  
Started by seqadmin, 07-15-2024, 06:53 AM
0 responses
34 views
0 likes
Last Post seqadmin  
Started by seqadmin, 07-10-2024, 07:30 AM
0 responses
41 views
0 likes
Last Post seqadmin  
Started by seqadmin, 07-03-2024, 09:45 AM
0 responses
205 views
0 likes
Last Post seqadmin  
Working...
X