Will Magnetic Beads clean & concentrate HMW circular plasmid DNA?

20 June 2017 0 7K Report

We are currently using PCR clean-up columns to purify 14Kb circular plasmid DNA & getting low yields in large volumes. We routinely use Magnetic Beads (AmpureXP or Kapa) to clean & concentrate NGS libraries, does anyone have experience using beads to clean & concentrate high molecular weight circular plasmid DNA? Thank-you in advance! Selene.

Badges
Science topic

Similar topics
Computer Science
Program

More Sélène Tyndale's questions See All

Is there an alternative to AraC to kill proliferative cells?

Hi researchgate community, I am planing to differentiate neurons from induced pluripotent stem cells and the protocol recommends to add AraC (Cytosine β-D-arabinofuranoside) at some point to kill...

02 April 2023 5,711 3 View

I have a protein tagged with MBP (maltose binding protein)with a TEV cleavage site between MBP and prot. How to improve the cleavage of the tag?

I have already tried different conditions (temperature, salt concentration, presence of reducing agents, different concentration of protease) but the removal of the tag is always very partial (...

05 December 2021 2,125 13 View

PCR looks good on gel, after purification, sequencing looks bad?

I performed a PCR on some samples that looked good and only produced 1 band on a gel. After PCR purification with a Qiagen kit following normal procedures, I sent the samples out for sequencing. I...

16 July 2020 3,836 6 View

Countries focus on Biological Research?

I'm sorry if we are supposed to ask only about specific research topics, but lately, I've been wanting to learn a new language that could also help me on my research field. I'm not sure which one...

22 June 2020 3,260 4 View

PCR-free WGS on Illumina, will standard Y-shaped adaptors work? or are linear adaptors needed?

We have decided to do PCR-free Whole Genome Sequencing on Illumina sequencers. Will standard Y-shaped adaptors work? or do we need special linear adaptors? Any advice is appreciated.

24 October 2018 3,489 0 View

Will Concanavalin A beads help precipitate nuclei from lymphocytes?

I am trying to do an ATACseq library prep on human lymphocytes and keep losing the majority of cell nuclei in aspiration steps. It was suggested I could try using Concanavalin A beads to help...

21 March 2018 3,094 5 View

What is the best way of labeling a short peptide without changing its structure too much?

I would like to label a short peptide (30 aa) with a fluorescent dye. I'd like to avoid amine modification, since I have 3 lysins and I am afraid that labeling all of them would change the...

19 July 2015 3,164 6 View

Finding the right PhD

How do I find the right PhD? I'm going crazy about finding a PhD where I can combine the study of marine viruses and their effects on ocean environments (biological pump and biogeochemistry). If...

01 January 1970 4,616 3 View

Completely drying glassware before dry cycle autoclaving.

I seem to recall it being necessary to completely dry glassware before autoclaving on dry-cycle. However, a peer of mine doesn't think this is necessary and puts still wet glassware to autoclave...

01 January 1970 2,835 3 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Does crude extraction using NaOH and Tris work well with Fungi?

I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you

08 August 2024 4,733 2 View

E.coli contamination in human RNA seq data ?

Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...

06 August 2024 807 3 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View

Where to find a gene list for CRISPRa/i library screening of regulatory factors that affect pathogenic Th17 differentiation in PBMC?

I want to perform a CRISPRa/i library screening for candidate regulatory factors that affect pathogenic Th17 differentiation in PBMC. But I am wondering how to define the size of gene library, so...

31 July 2024 2,150 0 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View