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Sequencing Library Preparation

The MBCL offers library construction services for Illumina and PacBio platforms.

The service provided by the Core includes conversion of DNA or RNA into ligated & indexed libraries, size-selection, and QC by Fragment Analyzer and/or any other means necessary.

The Illumina library prep ticket can be found here as a printable version or as a fillable pdf.**

Required Material for Illumina Libraries
Illumina Library Types: Required Material*:
Genomic DNA 2-3 μg
Amplicon 1 μg
cDNA 1 μg
total RNA 2-5 μg
small RNA 50-100 ng

*Submissions with less than the minimum requirement can be submitted after approval by the Core. Email Maria.Zapp@umassmed.edu to discuss your project.

Required Material for PacBio RNA Libraries
PacBio Library Types: Required Material:
total RNA for IsoSeq/cDNA library 1 - 2.5 μg

**PacBio library preps from DNA/cDNA starting material are carried out by the PacBio Core. All PacBio samples, regardless of source material, should be submitted through the PacBio Core. Email Maria.Zapp@umassmed.edu to discuss your project and/or request a ticket.

Sample Drop Off Locations:

1. LRB - 6th floor copy room
2. ASC - 8th floor room 2058
3. Biotech 2 - fridge or freezer in the hall outside of room 207.
4. 222 Maple Ave. Shrewsbury, MA, 01545 - Rose Gordon Building, 1st floor. Available weekdays from 8AM to 5PM  (Email us if you leave samples here).

Samples get picked up at 11:00 AM every Monday through Friday.

Tips for isolating material:

1.  Samples should be suspended in water or EB. Other buffers or any contaminants could create problems with library preparation and be carried over into sequencing. Please avoid the use of glycogen whenever possible.

2.  NanoDrops and similar instruments frequently read concentrations higher (sometimes ten-fold more) than other methods of determining concentration. This is because the NanoDrop detects everything, including contaminants, in the sample. The Core recommends the use of other methods such as Qubit or Fragment Analyzer, or careful attention paid to NanoDrop calibration, 260/280, and 260/230 readings.

3.  Some suggested kits for DNA/RNA isolation are: (((coming soon!)))