Sample Submission Instructions

General Requirements and Sample Submission Guidelines:
DNA Amount:
We recommend that 100 nanograms (100 ng) of DNA be sent for each PCR reaction. If submitting 100 ng per PCR reaction is not feasible, contact Applied Gene technical support. We can help you to calculate the minimum DNA amount for your project.
 
Recommended RNA Isolation Methods:

• Cells: Extract and Purify with Qiagen RNeasy Kit (with on-column DNase treatment step, Qiagen Cat. No. 74104)
• Tissues: Extract with TRIzol® from Invitrogen (Catalog Number 15596-026), followed by purification of RNA using Qiagen RNeasy Kit with DNase treatment.

Note:

• DNase treatment is an essential step for SYBR green based PCR experiment.  Without DNase treatment step, genomic DNA may be detected and cause false positive signals.
• Dissolve in RNase-free water (or 10 mM Tris, pH 8.0) at a concentration about 0.5 mg/ml.  Do not use DEPC treated water.  Report actual concentration and A260/A280 and A260/A230 ratios in the table. 

Recommended Reverse Transcription Methods:

• High Capacity cDNA Reverse Transcription Kit with RNase Inhibitor from Applied Biosystems (Part Number 4374966)
• High Capacity RNA-to-cDNA Kit from Applied Biosystems (Part Number 4387406)

Note:

• The cDNA concentration should be around 100 ng/ml. If the concentration is not feasible, contact Applied Gene technical support. We can help you to calculate the minimum DNA amount for your project.

Confirmation of RNA Sample Integrity
Provide picture of ethidium-bromide stained agarose gel or analysis report from Agilent 2100
BioAnalyzer. Demonstrate sharp, intact 28S and 18S rRNA bands or peaks. Please note that poor RNA quality will result in poor real-time PCR results at the customer’s cost.