Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • pbluescript
    replied
    You can use the ratio of AMPure beads to sample to select a specific size range without using a gel. Since the beads are in a PEG/salt solution, altering the ratio of beads/sample will solubilize different sizes of DNA fragments. For example, using 0.6X AMPure beads removes DNA <500bp and using 1.2X removes DNA <150bp. Using two different ratios of beads will allow you to select a certain size range of your library. Using a 0.6X ratio will cause binding of DNA fragments >500bp, but the smaller fragments will still remain in the supernatant. You can take that supernatant and add it to a higher concentration of AMPure beads and then take the DNA bound to the beads to get a size selection based on the ratios you used. With the 0.6X and 1.2X ratios, your library would range from 150-500bp. Altering the ratios by 0.1X steps will allow you to collect narrower size ranges. This type of size selection isn't extremely accurate so you should only use it if appropriate for your application. Be sure to test this out on some 100bp ladder or something similar since there is a little bit of lot-to-lot variation.

    Leave a comment:


  • polyhedron
    replied
    Originally posted by Zaag View Post
    Ampure has a protocol for 96 and 384 well plates. Link to protocol (PDF):

    http://www.beckmangenomics.com/docum...000387v001.pdf
    thanks. but that's not size selection like cutting a narrow band from the gel, but rather just DNA purification.

    Leave a comment:


  • josdegraaf
    replied
    For tighter sizes pippin or Caliper labchip XT although no plate format for those..

    Leave a comment:


  • Zaag
    replied
    Ampure has a protocol for 96 and 384 well plates. Link to protocol (PDF):

    As a leader in R&D genomics services, GENEWIZ provides superior data and high-quality constructs for next generation sequencing, gene synthesis, and sanger sequencing.

    Leave a comment:


  • polyhedron
    started a topic high throughput DNA size selection?

    high throughput DNA size selection?

    Hi! I'm asking is there any method out for a large quantity of DNA size selection, something like with 96-well plate. The traditional way is to excise agaroase gel and do DNA purification, but it's too labor-intensive for hundreds of samples. I think it would be perfect to do size selection in a 96-well plate. But something like Sephadex seems not able to do such selection (e.g. 250-300bp). Any idea? Thanks!

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, 07-10-2024, 07:30 AM
0 responses
23 views
0 likes
Last Post seqadmin  
Started by seqadmin, 07-03-2024, 09:45 AM
0 responses
198 views
0 likes
Last Post seqadmin  
Started by seqadmin, 07-03-2024, 08:54 AM
0 responses
209 views
0 likes
Last Post seqadmin  
Started by seqadmin, 07-02-2024, 03:00 PM
0 responses
191 views
0 likes
Last Post seqadmin  
Working...
X