BioMicroCenter:Multiplex: Difference between revisions
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[[IMAGE:BioMicroCenter_Barcode.png]] | [[IMAGE:BioMicroCenter_Barcode.png]] | ||
5' Barcodes have the advantage that they are read as part of a single read. The disadvantage of the barcode is they require two primers to make (both halves of the Y-shaped adapter) and, due to the way Illumina finds clusters, you must use a large number of barcodes in each run (16+). In addition, because they are part of the ligation event, care must be taken that the barcode is not introducing biases in library construction. 3' Barcodes are more flexible, being added during the PCR step, and can have a number of different compositions and can be run in any number. The do require a second read to sequence which can add extra cost to the flowcell. | |||
== MULTIPLEXING IN BIOMICRO == | == MULTIPLEXING IN BIOMICRO == | ||
Revision as of 11:40, 11 November 2010
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MULTIPLEXING
Multiplexing methods allow for running multiple samples in a single Illumina lane. There are multiple strategies for multiplexing Illumina samples but they can generally be broken down into two classes: 5' barcodes and 3' barcodes.
5' Barcodes have the advantage that they are read as part of a single read. The disadvantage of the barcode is they require two primers to make (both halves of the Y-shaped adapter) and, due to the way Illumina finds clusters, you must use a large number of barcodes in each run (16+). In addition, because they are part of the ligation event, care must be taken that the barcode is not introducing biases in library construction. 3' Barcodes are more flexible, being added during the PCR step, and can have a number of different compositions and can be run in any number. The do require a second read to sequence which can add extra cost to the flowcell.
MULTIPLEXING IN BIOMICRO
The BioMicro Center uses 3' barcoding for our sample preparation. Barcoding is available for *FREE* as part of our full Illumina sample preparation service and are added during the amplification step. The barcodes we use are 6nt in length and are separated by at least 2nt from any other barcode. We currently have a stock of 40 barcodes and have designs available for over 100.
Reading BMC barcodes requires a re-prime of the sequencer and there are some additional charges multiplexing samples. A full list of our prices is here.
