biogem: LightCycler 480 Roche QPCR Sample Submission Form

LightCycler 480 Roche QPCR Sample Submission Form

UNIVERSITY OF CALIFORNIA, SAN DIEGO
BIOMEDICAL GENOMICS MICROARRAY CORE FACILITY

RNA Sample Requirements for LightCycler 480 Assays

Please read this carefully.

This form must be filled out completely before we can accept any samples. Once this form has been filled out, please coordinate with Colleen Eckhardt to drop off samples.

A critical aspect of any experiment involving RNA is that the quality of that RNA and failed experiments due to problems with RNA quality is something that BIOGEM wishes to avoid. Therefore we require that the RNA is prepared with the RNeasy kit from Qiagen. This ensures high quality RNA that will not contain contaminants that interfere with enzymatic reactions.

In addition to the tubes of RNA supplied for cDNA synthesis, please provide a separate aliquot for Agilent Bioanalyzer analysis. Provide the RNA for Agilent analysis at a concentration range of 0.3 mg to 1 mg per ml at a volume of no less than 3ul. Label these tubes with an "A" (denoting they are for Agilent analysis).

Label the labeling aliquot tubes with these information:

Your name.
Date of sample submission (day/month/year).
Name of platform requested ("Roche 480"). Summary of RNA Requirements:
Amount of RNA REQUIRED for Agilent Bioanalyzer analysis: 0.3mg to 1 mg/ml at a volume of no less than 3ul. Do NOT dilute the Bioanalyzer aliquot as concentration of the labeling aliquot will be based on this.
Amount of RNA REQUIRED for cDNA synthesis: 1-2ug total RNA. (1 ug is needed to generate 20ul of cDNA, which is enough for 10 qPCR reactions. We keep the extra in case the cDNA fails the first time. We return any RNA we do not use if that is what the investigator wishes).
RNA 260/280 between 1.8-2.0.
Gel validation (if RNA quantities permit).
Ratio of high/low molecular weight rRNA: ~2.
RNA must be resuspended with RNAse-free water (NOT DEPC WATER) at a concentration within the range of 0.1 to 1 mg/ml, concentrations outside this range will incur an extra charge if we have to concentrate or dilute the material. Please let us know in advance if the RNA is not at 0.1-1 mg/ml.

RNA quality will be checked at the facility. If low quality is suspected or the RNA fails to meet our criteria, the experiment will be stopped. Please contact Colleen Eckhardt if you have any questions or concerns about the integrity of your RNA samples.

Fill out the form

Last Updated March 2008
By Dr. Gary Hardiman

LightCycler 480 Roche QPCR Sample Submission Form

UNIVERSITY OF CALIFORNIA, SAN DIEGO BIOMEDICAL GENOMICS MICROARRAY CORE FACILITY

RNA Sample Requirements for LightCycler 480 Assays

UNIVERSITY OF CALIFORNIA, SAN DIEGO
BIOMEDICAL GENOMICS MICROARRAY CORE FACILITY