Tissues & Cells

For many investigators, tissues or cells will be the major source of total RNA for expression profiling. Since these specimens contain nucleases, quick and careful collection of specimens is critical to ensure intact total RNA for analysis.

Expression Analysis requires 2 ug of total RNA (Affymetrix) and 100 ng of total RNA (Illumina) for our standard process; however, we can accommodate specimens with a minimum of 5 ng. To ensure that the isolation procedure will yield a sufficient quantity of total RNA, we recommend that you research the amount of tissue or cells required to yield the minimum quantity prior to harvesting your specimens. We recommend referring to the guidelines set for the QIAGEN RNeasy kit, which can be obtained at the QIAGEN website (www.qiagen.com).

For your convenience, we have provided the table of recommended tissue amounts below for the most popular tissue types.

Table 1. Recommended Tissue Amounts to Yield 5-10 ug of Total RNA.
Tissue	Size Range (mg)	Tissue	Size Range (mg)
Bone Marrow	35-60	Liver	20-30
Brain	60-80	Lung	50-70
Heart	23-35	Smooth Muscle (stomach or intestine)	35-50
Kidney	20-30

For best results, Expression Analysis recommends the following procedure for harvesting tissues:

Clean all instruments, bench tops, and gloves before starting with an appropriate RNase decontamination solution.
Harvest tissues as soon as possible after euthanasia and immediately freeze in liquid nitrogen. If liquid nitrogen is not available, you can use a dry ice/alcohol bath.
Weigh frozen tissue on a lab balance, and record quantity on the Expression Analysis Specimen Submission form.
If immediately freezing the specimens is not possible, place the tissue in an appropriate RNA stabilization reagent such as RNAlater™.
After collection, store specimens at -80ºC for shipment.
See Shipping Specimens section for shipping instructions.

Preparation of Cells

To ensure the extraction process will yield enough total RNA for analysis, Expression Analysis recommends that you send between 5 x 105 and 107 cells per specimen. For best results, Expression Analysis recommends the following procedure to prepare cells for shipment:

When a sufficient number of cells have been grown, pellet cells and pour off media.
Wash cells 2X with an appropriate volume of PBS, pellet cells, and remove all remaining PBS leaving only the cell pellet.
Do not send specimens in lysis buffer.
Snap freeze cells in liquid nitrogen or dry ice/alcohol bath.
Store specimens at -80ºC for shipment.
See Shipping Specimens section for shipping instructions.