PCR+conditions+that+worked+well+for+amplifying+some+chimeric+transcripts+7-2-13

PCR conditions that worked well for amplifying some chimeric transcripts 7-2-13 ^specifically the EATr (2667 bp) and RS8 (740 bp) transcripts were amplified

experiment performed 7-2-13

fresh polymerase and dNTPs were used

Reaction -1 uL 4T1 cDNA (4T1 Tumor 1st strand cDNA 11-30-12 KW) -22.5 uL H20 -1 uL 2 uM forward primer -1 uL 2 uM reverse primer -25 uL PrimeSTAR Max Premix (hot start. .so primer was not added until the denaturation high temperature was reached)

Thermocycler conditions: Thermocycle Conditions 7-2-13 Note that the first 30 cycle amplification probably could have been reduced to 20 cycles since the bands were so strong.

Scan of page from notebook with annotated gel image here "F:\kurt\storage\CIM Research Folder\DR\2013\7-2-13\scan of chimeric transcript pcr 7-2-13.pdf"