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NEXTflex™ BRCA1 and BRCA2 Amplicon Panel Illumina-Compatible (48 Barcodes) Bioo Scientific NOVA-4220-02 24 rxns £2768.00
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Product Detail: The NEXTflex BRCA1 & BRCA2 Amplicon Panel is a complete, validated solution that targets all exons in BRCA1 and BRCA2 loci using 130 PCR primer pairs.

This kit requires only 20 ng of DNA from fresh or frozen samples. The convenient predesigned panel allows researchers to focus on data generation and analysis, not on labor-intensive primer design and target selection. While competitor protocols typically require several hours to perform, the NEXTflex BRCA1 & BRCA2 Amplicon Panel produces targeted Illumina-compatible libraries in only 2 hours. Additionally, the NEXTflex BRCA1 & BRCA2 Amplicon Panel protocol can be easily automated on liquid handlers, further simplifying workflow.

[BENEFITS]
>>> Complete BRCA1 and BRCA2 coverage
>>> 100% uniformity of all targeted coding exons and exon-intron boundaries
>>> Shortest Illumina targeted library prep protocol available - 2 hour protocol validated with low input samples
>>> For detection of germline mutations multiplex up to 384 samples at 100x coverage and for detection of somatic mutations multiplex 14 samples at 2000x coverage on a single Illumina 2x300 MiSeq lane
>>> Complete solution for targeted sequencing including gene specific primers, PCR Master Mix, and up to 384 barcodes
>>> Low input - only 20 ng DNA required from fresh or frozen tissue or blood

The NEXTflex™ BRCA 1 & BRCA 2 Amplicon Panel contains enough material to prepare 8, 48 or 96 samples for Illumina® compatible sequencing from DNA isolated from fresh or frozen tissue. The shelf life of all reagents is 12 months when stored properly. All components can be safely stored at -20°C.

For larger volume requirements, customized and bulk packaging is available. Please email us to tech@nktscientific.com for further information.

This kit is not designed for use with DNA isolated from FFPE or cfDNA samples.

Product Type: NGS - Illumina Amplicon Panels
Applications: Illumina BRCA1 and BRCA2
Data Sheet: PLEASE CLICK HERE TO DOWNLOAD
Search Terms: Illumina BRCA1 and BRCA2
Additional Info: [Recommendations for the Analysis of Sequencing Data]

Sequencing:
We recommend paired-end sequencing on a MiSeq with 2 x 300 bp reads (v3 cartridge) or 2 x 150 or 2 x 250 bp reads (v2 cartridge). If the full capacity of the cartridge can be used, then 2 x 300 bp v3 cartridges is more economical option than 2 x 150 or 2 x 250 bp v2 cartridges because, while higher priced, 2 x 300 bp v3 cartridges provide the option to sequence twice as many reads compared to v2.

Adapter Trimming :
While the MiSeq software provides an adapter sequence masking option, we recommend using a dedicated trimmer software, to ensure complete 3' adapter removal. We recommend the Trimmomatic. This software is best used in paired end mode. The minimum length of reads not removed after adapter trimming should be set to 69 (which is the length of the longest possible primer dimer).

Primer Sequence Removal:
Primer sequences, which do not provide useful sequence information, can be removed from trimmed reads before the alignment. We suggest the use of Cutadapt software in paired 5' anchored adapter removal mode. This adapter removal mode will remove the 5’ primer sequences of all amplicons provided with the kit from both R1 and R2 reads. The primer fasta file should be modified by inclusion of a caret sign (^) at the 5' end of each primer sequence. 3' primer sequences which could be more difficult to unequivocally identify can be removed by instructing Cutadapt to remove 35 bp (which is the length of the longest primer) from the 3' end of each read. For 2 x 150 sequence results, do not remove the 3’ primer sequence.

Alignment:
Bowtie2 aligner in end-to-end alignment mode and other commonly used aligners (e.g. BWA) may be used. Using the paired reads alignment option is recommended. The BEDfiles provided are for the hg38 version of human genome.

Mutation Calling:
For mutation calling, SAM tools mpileup can be used, followed by bcftools, to produce binary bcf files which are then converted to vcf. Alternatively GATK Unified Genotyper of the GATK pipeline could be used.
Application Details: Please use our contact form above to request a copy of the datasheet
Catalogue Number Update: Please note, the catalogue number of this item has changed to include the prefix NOVA-. The original catalogue number (without the "NOVA-" prefix) will still be valid, and the item shipped will be the same as only the catalogue number has changed. The original catalogue number was 4220-02
NGS Applications: BRCA1 and BRCA2 Panels