PaxGene Blood RNA Kit Handbook - Plastisch chirurgen

Troubleshooting GuideThis troubleshooting guide may be helpful in explaining any questions that may arise.The scientists in QIAGEN Technical Services are always happy to answer any questionsyou may have about the information and protocol in this handbook (see page 34 forcontact information).Comments and suggestionsRNA degradedRNase contaminationBe careful not to introduce any RNases into thereagents during the procedure or later handling(see Appendix A, page 29).Low RNA yielda) Less than 2.5 ml blood Ensure that 2.5 ml blood is collected in thecollected in the PAXgene PAXgene Blood RNA Tube (see PAXgene BloodBlood RNA TubeRNA Tube Product Circular).b) RNA concentration measured RNA concentration must be measured in 10 mMin waterTris·Cl, pH 7.5 for accurate quantification(Appendix B, page 30).c) Cell debris transferred to the Avoid transferring large particles when pipettingPAXgene RNA spin column in the supernatant in step 7. (Transfer of smallsteps 9 and 10debris will not affect the procedure.)d) Supernatant not completely Ensure the entire supernatant is removed. If theremoved in step 3supernatant is decanted, remove drops from therim of the tube by dabbing onto a paper towel.Take appropriate precautions to prevent crosscontamination.e) After collection in the Incubate blood in the PAXgene Blood RNA TubePAXgene Blood RNA Tube, for at least 2 hours after collection.blood is incubated for lessthan 2 hoursPAXgene Blood RNA Kit Handbook 06/2005 27