GLP-compliant FAQs
Confirmatory Sanger Sequencing
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We accept PCR products (crude or purified), RNA, cDNA, cell pellets, fresh/frozen tissue samples, and viral particles.
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Yes, for confirmatory sequencing a reference sequence is required. If you do not want to share your reference sequence with us, GLP-grade Standalone Sanger Sequencing may fit your needs.
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Yes, we can design the assay(s) to target the region of interest based on the provided reference sequence.
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You will receive a GLP-grade study report reviewed by the Azenta Quality Assurance department which includes strategies used, consensus sequences, and any sequence discrepancies detected by comparison to the reference sequences. We will also provide raw sequencing data in .ab1, .phd, and .seq formats, a signed GLP Compliance Statement provided by the Study Director, and a signed Quality Assurance Statement listing project inspection and reporting dates.
SNP/Mutation Analysis
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SNP/Mutation Analysis is a PCR plus sanger sequencing technique targeted toward analyzing Single Nucleotide Polymorphisms (SNPs) and mutations at the genomic DNA level.
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We accept Purified gDNA, cell pellets, fresh/frozen tissue samples, yeast colonies, and bacterial colonies.
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Yes, we can design assays to target your region of interest. To do so, we request either the accession number, the chromosome position along with genome build, the name of the gene of interest, or SNP ID (rs#).
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You will receive a GLP-grade study report reviewed by the Azenta Quality Assurance department which includes strategies used, consensus sequences, and any sequence discrepancies detected by comparison to the reference sequences. We will also provide raw sequencing data in .ab1, .phd, and .seq formats, a signed GLP Compliance Statement provided by the Study Director, and a signed Quality Assurance Statement listing project inspection and reporting dates.
GLP-Grade Standalone Sanger Sequencing
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We accept Purified PCR products (Pre-Mixed or Pre-Defined reactions) and Purified Plasmid DNA (Pre-Mixed or Pre-Defined reactions).
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A “Pre-mixed” reaction is submitted with DNA at an adjusted sample concentration that either already contains sequencing primers from the customer, or will use a free Azenta universal primer. Azenta sample submission guidelines must be followed with this method .
A “Pre-defined” reaction is submitted with DNA at an adjusted sample concentration that does not contain any sequencing primers. You will need to supply your primer in a separate tube for Azenta to add to your reaction. Azenta sample submission guidelines must be followed with this method.
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No. For standalone sequencing, only material that can be directly sequenced can be submitted. If PCR is needed prior to sequencing, our Confirmatory Sanger Sequencing service line may fit your needs.
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You will receive a GLP-grade study report reviewed by the Azenta Quality Assurance department which includes quality scores of all sequencing traces, raw sequencing data in .ab1, .phd, and .seq format, a signed GLP Compliance Statement provided by the Study Director, and a signed Quality Assurance Statement listing project inspection and reporting dates.
Plasmid DNA Preparation
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To confirm plasmid identity after preparation, the purified plasmid will be analyzed through restriction digest and sanger sequencing.
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A GLP-grade study report reviewed by the Azenta Quality Assurance department which includes strategies used, consensus sequences, and any sequence discrepancies detected by comparison to the reference sequences. You will also receive raw sequencing data in .ab1, .phd, and .seq formats, a signed GLP Compliance Statement provided by the Study Director, and a signed Quality Assurance Statement listing project inspection and reporting dates. A custom certificate of analysis (CoA) will also be included with the plasmid preparation.
Nucleic Acid Extraction
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We accept cell pellets, fresh/frozen tissue samples, and viral particles.
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You will receive a GLP-grade study report reviewed by the Azenta Quality Assurance department, a signed GLP Compliance Statement provided by the Study Director, and a signed Quality Assurance Statement listing project inspection and reporting dates.
Have a specific question?
Email | Phone 1-877-GENEWIZ (436-3949), Ext. 3350
SNP/Mutation Analysis
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SNP/mutation analysis is a PCR and Sanger sequencing technique for analyzing single nucleotide polymorphisms (SNPs) and mutations in genomic DNA.
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We accept purified gDNA, cell pellets, fresh/frozen tissue samples, FFPE samples, blood, buccal swabs, saliva, and yeast and bacterial colonies. Please inquire about other sample types by adding a comment in the “comments section” of your order form or by sending an email to molgen@azenta.com.
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Yes, we can design assays to target your nucleotide region of interest. We request submission of the accession number or chromosome position, genome build, and the name of the gene or SNP ID of interest. You may also provide a reference sequence in text format with your target(s) of interest highlighted, and 200 bp of flanking sequence on either side of the target(s).
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Yes, you can send us assays originally designed in your lab. We will charge an assay optimization fee to ensure that the assay is robust and functional in our lab. You can either supply custom primers with your sample or give us the sequencing information and we will synthesize the primers.
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Amplicons are targeted to be 400-500 bp for standard samples and 150-250 bp for FFPE samples. However, we can design larger PCR amplicons for an additional cost. This may vary depending on the complexity of the region of interest for a particular project.
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The fixation procedure tends to introduce some degradation into formalin-fixed paraffin embedded (FFPE) samples. With FFPE samples, smaller amplicons are required to maximize success rates in the PCR assay because it is much more difficult to analyze degraded DNA containing large amplicons.
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Yes, we can but that will incur additional charges according to the size of the target region. Although standard sample processing involves end-sequencing, if you need additional coverage for the entire amplicon, please make the request in the “comments section” of the order form.
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You will receive the original sequencing results in a chromatogram (.ab1 files) and the sequence files (.seq files).
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Standard analysis includes mutation calls made at the nucleotide level. You may request “custom analysis” including translated amino acid calls for an additional fee.
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We will process the samples once with the expectation that 80% of the sequences will be deemed “clean.” In case we do not attain that threshold, we will automatically repeat the assay free of charge.
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If it is necessary, we will perform a single repetition of the PCR and sequencing reactions in order to achieve cleaner traces.
16S rRNA Gene Sequencing
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This is a PCR and Sanger sequencing technique for identifying bacterial strains through the analysis of the 16S ribosomal RNA (16S rRNA) gene. It contains highly conserved regions between different species of bacteria, and this allows for targeting via PCR. After data analysis, the differences between species identified through Sanger sequencing, allows you to distinguish between individual samples.
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We accept clonal colonies on agar plates, glycerol stocks or purified gDNA samples.
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We use a Universal Primer set for the assay but the actual sequences are proprietary.
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The primers target the V1 to the V9 regions of the 16S rRNA gene.
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The PCR assay generates a 1.5kb PCR product.
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No, this service is for clonal samples that are not from a mixed population.
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You will receive the original sequencing results in a chromatogram (.ab1 files) and the sequence files (.seq files).
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No, but you can use the raw data we provide to identify bacterial strains.
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No, this service is not applicable to yeast.
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Although we have processed 18S assays submitted by our customers, currently we do not offer a universal 18S in-house assay.
cDNA Verification
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We accept RNA, cDNA, cell pellets, fresh/frozen tissue samples, FFPE samples, blood, and viral particles. Please inquire about other sample types by adding a comment in the “comments section” or by sending an email pm@azenta.com.
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Yes, we can design assays to target your region of interest. We request submission of information pertaining to the transcript(s) of interest per gene. If multiple transcripts need to be targeted, please include that information in your quote request and the project will be designed accordingly.
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Yes, we can design assays to amplify endogenous and recombinant targets. For endogenous targets, we request submission of information pertaining to the transcript(s) of interest per gene. For recombinant targets, please submit the reference sequence for the target of interest, including any flanking sequence surrounding the target, at least 100 bp 5’ and 3’ of the target region, if available.
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Yes, previously designed assays can be sent to the lab. The lab will need to ensure that the assay is robust and functional in our lab. Primers can either be submitted with the samples or the primer sequence information can be submitted and we will order the corresponding primers.
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We will design the primers based on the 5’ and 3’ end of the reference sequence you submit. Please note that the primers will mask the natural sequence on these ends of the reference sequence.
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No, we do not offer genome walking services. We can verify that the region of interest exists, but we will not go beyond that to identify the gene location with this method.
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Yes, we include two controls (i.e., no reverse transcriptase and no template controls) during the reverse transcription reaction.
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We will send you the original sequencing results in chromatogram format (.ab1 files), the sequence files per trace (.seq files), a consensus file of the aligned traces (.seq), and an alignment file of the consensus sequence against the reference sequence (.pdf).
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A custom report includes the primers used in the project and a consensus sequence for the region of interest. Any discrepancies between the sequence traces and the reference sequence will be highlighted within the report.
gDNA Verification
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gDNA verification is a PCR and Sanger sequencing technique for confirming longer stretches of a sequence in a gDNA sample. The gDNA is used as template for PCR in order to target the region of interest. Sanger sequencing is then used to confirm the sequences in the target region. Typically, this application is used to confirm changes made during cell line engineering experiments.
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We accept gDNA, cell pellets, fresh/frozen tissue samples, FFPE samples, blood, and viral particles. Please inquire about other sample types by adding a comment in the “comments section” or by sending an email to pm@azenta.com.
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Yes, we can design assays to target your region of interest. We request submission of information pertaining to the particular gene of interest or region of gDNA. If an engineered sequence needs to be targeted, please submit this sequence information in your quote request and the project will be designed accordingly.
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Yes, we can design assays to amplify recombinant targets. For recombinant targets, please submit the reference sequence for the target of interest including flanking sequence surrounding the target, at least 100 bp 5’ and 3’ of the target region, if available.
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Yes, previously designed assays can be sent to the lab. The lab will need to ensure that the assay is robust and functional in our lab. Primers can either be submitted with the samples or the primer sequence information can be submitted and we will order the corresponding primers.
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We will design the primers based on the 5’ and 3’ end of the reference sequence you submit. Please note that the primers will mask the natural sequence on these ends of the reference sequence.
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No, we do not offer genome walking services. We can verify that the region of interest exists, but we will not go beyond that to identify the gene location with this method.
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We will send you the original sequencing results in chromatogram format (.ab1 files), the sequence files per trace (.seq files), a consensus file of the aligned traces (.seq), and an alignment file of the consensus sequence against the reference sequence (.pdf).
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A custom report includes the primers used in the project, consensus sequence for the region of interest, and a highlight of any discrepancies between the sequence traces and the reference sequence.