Hey, bought a Roche lightcycler 2 system second hand :- I also have no idea where to get the software from it :- can't affort the $5000 version 4.1 that Roche demands online which is considerably more than I paid for the machine itself:- anyone willing to sell me a CD of an older or current version? Ta.
Samples were incubated for 60 min at 42C in a thermocycler. Real-time PCR was performed in duplicate with LightCycler 480 SYBR Green I Master reaction mix (Roche, Mannheim, Germany) and a LightCycler 480 system (Roche). PCR reactions were performed with 1 μl cDNA, 5 μl primer mix (containing forward and reverse primer, 500 nmol/l each), 10 μl 2x concentrated LightCycler 480 SYBR Green I Master mix (Roche), and 4 μl nuclease-free water. Relative mRNA levels were determined by using standard curves for each gene, and quantitative normalization of gene expression was performed in relation to the expression of the housekeeping gene RPS9. As a template for the standard curves, a mixture of all samples in 4 serial dilutions was used, corresponding to 3, 1, 0.1, and 0.01 μl template amount. The following PCR temperature cycles were used (45 cycles were performed): Step 1: 95C, 5 min; Step 2: 95C, 10 sec; Step 3: 55-60C (primer dependent), 10 sec; Step 4: 72C, 10 sec; Step 5: 4C, hold. Real-time PCR data were analyzed with LightCycler 480 software (Roche).
lightcycler 480 software free download
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