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Long PCRSeq

 
 
Explore expanded possibilities with Microsynth's Long PCRSeq, leveraging the cutting-edge long-read sequencing technology from Oxford Nanopore Technologies (ONT) to sequence clonal linear DNA ranging from 600 bp to 50 kb in length. Conveniently accessible for samples in tubes and 96-well plates, this service builds upon the capabilities of our renowned Full PlasmidSeq.
 

Features and Benefits


Fast
  • Results delivered within 1 to 2 days of sample receipt
  • Results are delivered Monday to Friday
 
Convenient
  • Use our drop box in your vicinity for free shipping of samples.
  • You can choose between prepaid and non-prepaid service.
 
Easy
  • Hypothesis-free sequencing of the complete PCR product.
  • No primers needed.
 

Cost-Effective
  • Economical way of sequencing your PCR product (amplicon).
 
Complete Information
  • You will receive the polished consensus sequence in different formats (e.g., fasta, fastq and ABI)
  • The annotation of the consensus sequence is available as html and genbank files.
  • The thorough uncertainty analysis of the consensus provides detailed information on sequencing and assembly quality of your sample.
 

FAQ

Logistics

How can I ship the samples?

Use our drop box system, or send your samples by mail.

What amount of DNA and concentrations do you need?

>15 µl at 20 ng/µl

Technical Questions

How can I judge the quality of the received sequence?


We supply additional information that allow you to identify problematic regions and evaluate the overall sequencing quality:

For instance, in the *.uncertain.tsv file that opens with Excel, all residues of the contig are listed that show a rate >25% of mismatches, deletions and/or insertions. These values are based on all the reads that were used to create the contig. You may sort the list according to % mismatches, % deletions, or % insertions to identify the regions within your plasmid that might be problematic or ambiguous. The coverage, as well as base quality of the reads at the respective position are also indicated.
In the *.clean.reads.html file of the Sequencing subfolder of your package you can inspect the statistics of the reads that were used to assemble your sample. The statistics comprise number of reads, length of reads and quality score of reads.

How can I open/view the data in certain files (eg. tsv, gbk)?

If you are working on Windows you may want to switch on viewing of file name extensions (In File Explorer under View, in the Show/hide group, select the File name extensions check box). For instance tab separated value files (.tsv) may be opened in any text editor or by drag and drop in an open but empty Excel table. Fasta (.fasta) and genbank (.gbk) files may be opened in a text editor or by specialized software.

Why are some terminal nucleotides missing from my linear/amplicon consensus sequence?

Depending on the sequencing chemistry used, the assembler may face challenges reconstructing the terminal ends of linear DNA, potentially leading to the absence of up to ~25 nucleotides at the 3’ and/or 5’ ends of your insert.

How to Order

 
 

The Long PCRSeq can be ordered through the same forms and labels as the Full PlasmidSeq.

Procedure to order labels for our Full PlasmidSeq/Long PCRSeq:

  • Enter our webshop
  • Click on Full PlasmidSeq in the "Order Labels" area and follow the further instructions
  • You can choose between prepaid and non-prepaid labels
 
Procedure to use our Full PlasmidSeq for sequencing.

Tubes:
  • Enter our webshop
  • Click on Fill Order Form under Full PlasmidSeq (prepaid or non-prepaid) and follow the further instructions
Plates:
  • Enter our webshop
  • Click on Fill Order Form under Full PlasmidSeq and follow the further instructions
 
See also Microsynth's user guide on the right side for more detailed information.