Optimization of RNA Extraction Achieved!

 Today is the first day of Ramadhan 1442 Hijrah where Muslims start to fast and so do I. So today, I came to the lab around 10 am to prepare for the final optimization steps of RNA extraction from C. lyoni spiderlings. After preparation, I started working at 1130 am and finished at 130 pm and finally managed to analyze the RNA bands on the bleach gel. I found that incubation of the RNA with formalaldehyde and heated for 2 minutes at 70 degree celcius has the same effect as at 65 degree celcius for 15 minutes. Incubation at 65 degrees for 5 minutes did not denature well the RNA secondary structure. Therefore, I opted to use pretreatment with aldehyde at 70 degrees for 2 minutes as the optimum point. From this experiment, the 28S rRNA is about 5000 bp while the 18S rRNA is approximately 1800 bp. Note that the intensity ration of 2:1 for 28S and 18S does not apply to invertebrates. 

                                    At 70 degrees                                                      At 65 degrees