Sample Preparation

Before sequencing, nucleic acid samples must be converted into libraries of short DNA fragments ligated with adapters which bind the DNA to the sequencing flowcell. Researchers may prepare their own libraries or the core can prepare them as a service. We can sequence libraries prepared with Illumina's kits, kits from third-party vendors, and custom libraries. If your library is non-standard, please let us know.

Automated Library Preparation: To request full-service library preparation, use the links below to download and complete the appropriate submission form and email it to illumina_submission-list@lists.fas.harvard.edu. Submissions which do not meet the input specifications listed on the form will be performed without guarantee. Our standard turn-around time is 6 weeks for "queued" library preps in which we combine samples from multiple users into one run. "Rush" preparation with a 3-week turn-around time is available when we have capacity - please inquire about the status of our queue.

DNA Preps

Input ng Chemistry Sample #
Kapa DNA Hyper Plus 1/4 Volume 12 - 122 ligation, incl. enzymatic fragmentation multiples of 8
Nextera XT 1/4 volume 0.05 - 0.25 tagmentation multiples of 8
RNA Preps Input ng Chemistry Sample #
Kapa Stranded RNA Hyperprep 50 - 500 ng stranded, incl. polyA selection multiples of 8
Kapa Stranded RNA Hyperprep with Riboerase 50 - 500 ng stranded, incl. ribosomal depletion multiples of 8, minimum of 24 samples
SmartSeq v4 1/4 volume 1 cell - 2.5 ng generates full length cDNA multiples of 8
Shearing: Library preparation involves fragmenting nucleic acid samples. Some protocols use enzymatic fragmentation, while some suggest mechanical shearing. For mechanical shearing, we recommend the Covaris S220 system which uses adaptive focused acoustics. Researchers may be trained to use the Covaris system independently. To request Covaris shearing service please email this form to illumina_submission-list@lists.fas.harvard.edu.

Size Selection: After library preparation, you must verify that your library is the optimal size for your downstream analysis and remove unwanted material such as adapter. Often, a bead-based size selection is adequate. For more precise size selection, we recommend the Pippin Prep and Blue Pippin systems which use special gel casettes available in a variety of agarose concentrations. Researchers may be trained to use both of these systems independently, or we can run them as a service.

Quality Control: Before submitting samples, we strongly recommend that researachers perform quality control checks on their libraries. We perform TapeStation and QPCR on libraries which we prepare. Researchers can elect to save on library preparation costs by performing QC on pools instead of on individual libraries. Please consult with the core and we'll help you select the appropriate level of QC for your needs.