Do libraries constructed with the Ion Xpress Barcode adapters require balancing to help ensure equal representation on the Ion PGM or Proton instrument sequencing run?
No, the Ion Xpress Barcode adapters are designed with a 3 bp adapter sequence (GAT) at the 3' end of each barcode to help minimize adapter ligation bias. When pooling barcoded libraries, accurate library quantification is extremely important to help ensure even barcode representation on the sequencing run.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How many fragment or amplicon libraries can be made per barcode with the Ion Xpress Barcode Adapters kits?
The Ion Xpress Barcode Adapters kits are available as a set of 96 (Cat. No. 4474517) or in sets of 16 (Cat. Nos.4471250, 4474009, 4474518, 4474519, 4474520 or 4474521). Each barcode kit is sufficient for preparing ? 10 libraries per barcode (10 x 16 libraries or 10 x 96 libraries) for 100 ng input, or 2 libraries per barcode for 1 µg input.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How many samples can I multiplex on a single chip for sequencing on the Ion PGM or Proton System?
The number of samples that can be multiplexed in a single sequencing run depends on the capacity of the chip, the size of the library, and the required coverage. A table of approximate capacities based on the size of the library and chip can be found in the Ion Ampliseq Preparation User Guide (https://tools.thermofisher.com/content/sfs/manuals/MAN0006735_AmpliSeq_DNA_RNA_LibPrep_UG.pdf) within the section “Strategies for combining Ion Ampliseq libraries.”
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
What are the differences between the Tag Sequencing Barcode Set 1-24 (Cat. No. A31830) and the Ion Xpress Barcode Adapters (Cat. No. 4474517)?
The TAG Sequencing Barcode Set contains adapters designed for Oncomine workflows using AmpliSeq HD technology and they can be used only with those library kits. This type of chemistry uses a molecular tag in order to track polymerase errors and increase the assay sensitivity. The Ion Xpress Barcode Adapters are not an alternative of the TAG Sequencing Barcode Set and vice versa. The list of the barcodes set compatible with a library kit is specified in the library manual.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How many samples from the Oncomine Comprehensive Assay Plus, manual library preparation can I run on the Ion 550 Chip?
We recommend sequencing up to 5 DNA or RNA samples (4 samples and a non-template control (NTC)) on a single Ion 550 Chip.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
