One of the most important criteria of next-generation sequencing library preparation is to start with high-quality, high-molecular-weight genomic DNA. This is especially true for long read sequencing on the PacBio Sequel.
Determining the quality of high molecular weight DNA can be done using pulsed-field gel electrophoresis. Pulsed-field gels are more effective than regular gels at achieving differential separation of the small and large DNA molecules due to the constant change in the direction of the electric field in the gel. Pulsed-field gel electrophoresis can resolve DNA out to 100 kb and beyond.
Please submit 50-100 ng of DNA in a maximum volume of 10 µl of 10 mM Tris buffer PH 7.5 or PH8 or in the Elution buffer (Buffer EB) from the Qiagen kit. Alternatively, DNA embedded in an agarose plug (CHEF Genomic DNA Plug kits from Bio-Rad) made with low-melt agarose can also be submitted, though liquid samples are preferred. Up to 18 samples and two lambda DNA size standards ranging from 5 kb to 430 kb can be loaded per gel. The lambda DNA size standards will be supplied by GGBC. The DNA will be run on a 0.75% agarose gel using the Pippin pulse electrophoresis power supply (Sage Science) overnight before being stained with the SYBR Safe DNA gel stain (Life Technologies).
A gel image showing resolution of the DNA fragments from the gel run with the Pippin Pulse will be provided to the customer.
Contact for Financial Inquiries and Quote Requests
Please email Kim and Elizabeth at firstname.lastname@example.org, for financial inquiries or to request a quote. Be as specific as possible, so that they can more quickly assist you.
Price per pulse-field gel up to 18 samples (includes two DNA size standard lanes per gel)
Table 1. Pulsed-Field Gel Electrophoresis
The price of the run can be prorated if customers share a run. Otherwise, the full price as advertised above will be charged regardless of the number of samples submitted.
Please note that there will be additional charges to quantify your sample using a Qubit assay before running the gel. PFGE is sensitive to the mass of DNA loaded, so an accurate concentration is essential to produce a clear gel image.