Genomics Application Notes

Reagents: • 100% Isopropanol; American Bioanalytical AB07015 or equivalent • 85% ethanol; freshly prepared/diluted from 100% ethanol (American Bioanalytical, AB00138 or equivalent)

• DNase I (RNase-free); Ambion AM2222 or AM2224 • Reagent grade water, nuclease-free; Ambion AM9932

Materials Supplied in the Kit

Storage Conditions once In Use (isopropanol, PK or buffer added)

Refered to in the Protocol As

Storage Conditions on Arrival

Reagent

Lysis Buffer

Lysis Buffer

Room Temperature

Room Temperature

Binding Buffer

Bind Buffer

4°C

4°C

Wash Buffer

Wash Buffer

Room Temperature

Room Temperature

Proteinase K

PK

-20°C

-20°C

Proteinase K Storage Buffer

PK Buffer

Room Temperature

-20°C

Kit Specifications The Agencourt RNAdvance Tissue kit is manufactured under RNase-free conditions and has been tested and certified not to contain contaminating nucleases. The Agencourt RNAdvance Tissue kit can be used in 96 well and single tube formats. General Remarks on Handling RNA RNases are ubiquitous and general precautions should be followed in order to avoid the introduction of contaminating nucleases during the Agencourt RNAdvance Tissue procedure. The most common sources of RNase contamination are hands, dust particles, and contaminated laboratory instruments, solutions and glassware. When working with RNA, the following procedures should be followed to limit RNase contamination: • Always work with gloved hands and change gloves frequently. • Use RNase free, filtered pipette tips for pipetting whenever possible. • Use dedicated RNase free equipment, e.g. pipettes, pipette tips, gels boxes, etc. • Avoid using reagents, consumables and equipment that are in common use for other general lab processes. • When available, work in a separate room, fume hood or lab space. • Use plastic, disposable consumables that are certified RNase free. • Purchase reagents, such as commonly used buffers and water, that are certified RNase free. • Prepare small individual aliquots of such buffers to avoid repeated transfer out of stock buffers. This lowers the risk of contaminating the stock solution. • Wipe down work surfaces with commercial RNase inhibiting surfactant solutions or 85% ethanol before starting work. Before starting • Preheat water baths or heat blocks to 37°C • Ensure that Agencourt RNAdvance Tissue Proteinase K and Wash Buffer have been assembled according to the following instructions: Agencourt RNAdvance Tissue 96 Well and Tube Procedure Assembly Steps 1 and 2 are only performed once for each new Agencourt RNAdvance Tissue kit. If these have already been completed during a previous experiment, please skip ahead to Step 3.

AAG-230APP07.14-A