SAFETY: One of the kit components contains the following chemicals:
8-10% of human urine (potential biohazard)
0.1-0.3% bovine serum albumin (harmful)
≤0.09% sodium azid (very toxic).
As both the amount and the concentration of the very toxic compound (sodium azid) is very low, the risk is also low.
Avoid contact with eyes, skin and other mucous membranes. Wash thoroughly after handling. Avoid breathing vapor/mist/spray.
Controls: Run a negative DNase treatment sample for every batch of DNase treatment to control for potential contaminations of reagents or during the treatment procedure.
Kit properties: Always write down the LOT number of the kit you use. Avoid using expired kits or reagents.
Test PCR for treatment: always perform test PCR (step 10-11.) to confirm the success of the DNase treatment.
VERY IMPORTANT: RNA is much more sensitive to degradation than DNA, therefore contamination with RNase has to be avoided (the main source is your own skin) and the samples have to be kept on ice all the time to slow degradation. The following steps help avoid degradation before and during RNA work:
Before starting to work clean bench, pipettes and the racks with MQ water (against DNA and RNA contamination), 70% ethanol (desinfection) and RNaseAway spray.
Use only nuclease-free filter tips and tubes.
Keep thawed RNA samples on ice all the time.
Use new gloves to touch samples and RNase-free pipettes and racks (cleaned in point 1). If you touch anything else (for example freezer handle), immediately change gloves. Change gloves regularly anyway during work with RNA, at least after every few steps. Before starting to work, spray RnaseAway on your gloves.
Special care should be applied until RNA is completely transcribed to DNA (end of RT process).