PCR-EZ Max Frequently Asked Questions

Samples may be submitted at any time. Samples must be received in the morning at our facility to be eligible for that day's cycle. Projects with incomplete information may be placed in the next sequencing cycle. Depending on your location, results may be delivered the same day or within 1-3 business days.

• Purified Linear PCR:

  • Sample Type: Purified Amplicon DNA

  • Size: 500 bp - 25 kb; if larger than 25kb, please email DNAseq@azenta.com for a technical consultation

  • Minimum Amount: 10 µL

  • Concentration: 25ng/µL per 1kb (i.e., for a 2 kb amplicon, provide 50ng/µLin 10 µL)

  • Purity (A260/280): 1.8-2.0 and (260/230): 2.0-2.2

  • Buffer: Water, EB, or low TE (<0.1 mM EDTA)
    
    

• Unpurified Linear PCR:

  • Sample Type: Purified Amplicon DNA

  • Size: 500 bp - 25 kb; if larger than 25 kb, please email DNAseq@azenta.com for a technical consultation

  • We recommend providing a gel image to help us estimate DNA concentration and optimize the dilution after enzymatic cleanup

  • Buffer: Water, EB, or low TE (<0.1 mM EDTA)

Submission Methods:  

• Drop off samples in a GENEWIZ dropbox at room temperature 
  
• Ship at room temperature or on blue ice
  

Tubes for <48 Samples​ 

• If you are submitting <48 samples, please use 8-strip, 0.2 ml PCR tubes and caps. You can cut off empty tubes and use them next time.​ 
• Our online ordering system will assign tube ID codes using your initials and the sample number (see figure below). Please write these codes on the sides of your tubes using indelible marker.​ 
• Label the side of your tubes with YOUR initials and sample number. These should match your order receipt.​
  

Plates for >48 Samples​ 

• If you are submitting 48 or more samples, we recommend using a 96-well, semi-skirted PCR plate. Cap the wells with 8-strip caps. These are usually ordered separately from the plates. Be sure that the caps seal tightly! The semi-skirted plate helps to prevent the plate from bending in transit, resulting in fewer loose caps. Please ship the plate with cushioning to avoid potential damage.​ 
• Please contact us for help finding a vendor if your laboratory does not have any 96-well PCR plates or matching strip caps.​ 
• Arrange your samples vertically (in columns) as shown below: 
  

Please send your samples with a copy of your order receipt to us using one of the following options.

Option #1: Submit samples into a local dropbox at room temperature. We have an extensive network of convenient dropboxes located throughout the US. To find a dropbox near you, please contact DNAseq@azenta.com. Place the samples and order receipt together in a Ziploc bag. 

Option #2: Ship samples directly to our facility using the shipping address listed on your order receipt. You can ship at room temperature or on blue/dry ice.

When submitting an order through your GENEWIZ account, the final step will locate the nearest dropboxes to your institution. You can then choose the dropbox where you will leave your sample for PCR-EZ Max. During checkout, consult the Order Summary page to find out the daily cutoff times for your selected dropbox (see example below).

If you miss the dropbox deadline, feel free to ship samples directly to us.

• Choose PCR-EZ for clonal, single-amplicon sequencing applications and characterization of up to 25 kb amplicons. Rapid barcoding is utilized, which can cause some fragmentation.
  
  

• Choose PCR-EZ MAX for mixed amplicon samples, low-frequency variant detection, and applications requiring higher sequencing coverage. Native barcoding ensures end-to-end coverage.

You can order PCR-EZ Max in minutes without a quote. Visit genewiz.com and log in to your online account to place an order.

The target coverage for PCR-EZ Max is 5000 reads per sample. However, the number of reads is relative to the number of amplicons and the size of the plasmids. Expect up to 5000 reads for <6 kb amplicons. 

The estimated delivery date for your order can be viewed anytime through your online account on the Order Summary page.

You will receive all the raw data reads for your sample. You also have the option to choose bioinformatic analysis (additional charges apply). 

Option 1: Project report, sample report, and raw FASTQ files

Option 2: Project report, sample report, raw FASTQ files, consensus assembly (FASTQ, FASTA, and ab1 formats), and variant report

Once data analysis is complete, your data will be delivered electronically through your online account. Please feel free to contact us should you have trouble retrieving your data.  

Yes, we take data security very seriously and make every attempt to keep your data private and protected. The data transfer we offer is secure. However, should you desire an alternative delivery method, we are happy to work with you.

All custom bioinformatic analysis requests are taken on a case-by-case basis. Please contact DNAseq@azenta.com  for a technical consultation. Please note that any customization outside our standard deliverables may incur additional charges and time.

It is likely that sequencing failure (i.e., failure of the sample to produce a consensus sequence with at least 10x coverage or less) may be due to samples not being prepared at the required DNA concentration or to degraded/fragmented DNA. 

To achieve optimal sequencing results, please follow our recommended sample submission guidelines.

We cannot guarantee the results of the sequencing data. If you feel that a processing error may have occurred, please contact us, and we will investigate on a case-by-case basis.