Amplicon-Seq

Amplicon-Seq

12.001,474.00

Amplicon-seq is an easy and inexpensive method for preparing focused NGS libraries from gDNA  for 1-5 targets. Amplicon-seq utilizes a first PCR process to generate the amplicon that will be the focus of the sequencing and a second PCR process to attach barcodes required for the NGS.

The first step PCR can be performed in your lab or at Syntezza.

The second step PCR is performed at Syntezza and then sent for NGS sequencing. When sequencing is competed, we can also facilitating bioinformatic analysis of the results.

 

SKU: N/A Category:
Image Variation SKU Price Quantity Add To Cart
Extraction 1-24 samples Amp-S Extract-1-24 40.00
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Extraction 25-48 samples Amp-S Extract 25-48 35.00
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Extraction 49-96 samples Amp-S Extract 49-96 30.00
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Extraction > 96 samples Amp-S Extract >96 25.00
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Both PCR Prep Steps 1-24 Samples Amp Prep Both 1-24 90.00
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Both PCR Prep Steps 25-48 Samples Amp Prep Both 25-48 85.00
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Both PCR Prep Steps 49-96 Samples Amp Prep Both 49-96 80.00
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Both PCR Prep Steps >96 Samples Amp Prep Both >96 75.00
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Barcoding Prep Only 1-24 Samples Amp Prep Barcode Only 1-24 60.00
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Barcoding Prep Only 25-48 Samples Amp Prep Barcode Only 25-48 55.00
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Barcoding Prep Only 49-96 Samples Amp Prep Barcode Only 49-96 50.00
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Barcoding Prep Only >96 Samples Amp Prep Barcode Only >96 45.00
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Sequencing 2x150bp 100K reads Amp Sequencing 2x150 100K 144.00
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Sequencing 2x250bp 100K reads Amp Sequencing 2x250 100K 221.00
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Sequencing 2x300bp 1M reads Amp Sequencing 2x300 100K 221.00
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Sequencing 2x150bp 1M reads Amp Sequencing 2x150 1M 961.00
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Sequencing 2x250bp 1M reads Amp Sequencing 2x250 1M 1,374.00
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Sequencing 2x300bp 1M reads Amp Sequencing 2x300 1M 1,474.00
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Ship GenTegra Amp-Seq Ship 12.00
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Description

gDNA extraction

Prior to preparing the amplicon library, we can pick up your  samples and perform the DNA  extraction at our facility or you can isolate the gDNA at your lab. DNA extraction service is available from ₪25.00 to ₪40.00 per sample.

Preparation of the amplicon library

The first step PCR reaction utilizes custom PCR primers designed to amplify your specific region of interest plus an additional 20 bp region upstream of each primer, called Linkers.

The linker sequence for the forward primer is CS1: 5’-ACACTGACGACATGGTTCTACA-[locus specific sequence – Forward primer].

The linker sequence for your reverse primer is CS2: 5’-TACGGTAGCAGAGACTTGGTCT-[locus specific sequence – Reverse Primer]

You can order the primers from IDT and perform the first stage PCR at your lab and simply send us the PCR product amplicons. We will then perform the second step PCR that adds a barcode to each sample to finalize your library in preparation for NGS sequencing. Pricing for this second step PCR is available from ₪45 to ₪60 per sample.

Alternatively, you can email your custom primer sequences to Syntezza. We will order the primers with the added linker sequences and perform the first step PCR as well as the second step PCR to finalize your library in preparation for NGS sequencing. Price for the complete Amplicon-Seq library preparation from gDNA is available for the cost of the custom primers plus ₪75 to ₪90 per sample.

The above processes are effective for NGS sequencing of at most a few PCR product targets in each sample well.

Amplicon library preparation for larger pools

An alternative system is required for larger amplicon pools to mitigate primer-dimer formation and proliferation of off-target PCR reactions. We offer cost effective methods for development of custom reagent sets that enable highly accurate and scalable NGS sequencing for dozens to hundreds of thousands of targets. Learn More.

Shipping

GenTegra technology enables shipping of DNA samples at ambient temperature without any fear of degradation, sample loss or delay in transport. Syntezza will deliver to your lab the appropriate number of GenTegra tubes required to stabilize your samples for transport to our facilities in Jerusalem and will use GenTegra tubes to safely transport your prepared libraries to the NGS sequencing facility. The cost of shipping and handling is a minimum ₪ 200 for the order, which includes GenTegra DNA tubes for up to 16 samples.

NGS sequencing

For Amplicon-seq libraries, we offer four sequencing options using Illumina MiniSeq or MiSeq instruments.

Number of Reads Read Size  Price
100,000 2 x 150 bp 144 ₪
100,000 2 x 250 bp 221 ₪
100,000 2 x 300 bp  221 ₪
1 M 2 x 150 bp 961 ₪
1 M 2 x 250 bp 1374 ₪
1 M 2 x 300 bp 1474 ₪

Our NGS specialists will help you select the best options and prepare a custom price quotation. Contact us today at NGS@syntezza.com or call 02-586-7138 ext. 2.

Additional information

Unit

Extraction 1-24 samples, Extraction 25-48 samples, Extraction 49-96 samples, Extraction > 96 samples, Both PCR Prep Steps 1-24 Samples, Both PCR Prep Steps 25-48 Samples, Both PCR Prep Steps 49-96 Samples, Both PCR Prep Steps >96 Samples, Barcoding Prep Only 1-24 Samples, Barcoding Prep Only 25-48 Samples, Barcoding Prep Only 49-96 Samples, Barcoding Prep Only >96 Samples, Sequencing
2x150bp 100K reads, Sequencing 2x250bp 100K reads, Sequencing 2x300bp 100k reads, Sequencing 2x150bp 1M reads, Sequencing 2x250bp 1M reads, Sequencing 2x300bp 1M reads, Ship GenTegra

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