Difficult sequencing

Early last week I received my first sequences back from Macrogen (a Korean sequencing biotech company) from the Multiplex trials I was running. When there is a lot riding on the results, I hate that period where you are just about to have a look at the document, knowing that if it is bad your working life is going to be a lot more stressful for the next week (or more…). I looked at the sequence chromatograms and, unfortunately for me, ¬†my sequences were very messy and thus difficult to interpret. Some sections were really good, but others were just plain ugly. How come I had really good bands but rubbish sequences? i was thinking I would have to go through the purification trial process all over again. Not a good start to my Monday morning. I was just about to drown my sorrows, when I got an email from Macrogen saying that they would do a retrial of some of the sequences for me. This flicker of hope kept me at the computer to do some research about what could have possibly gone wrong. After looking through the literature it looked like it could be the secondary structure problem all over again. Luckily Macrogen have dealt with this problem before, so I ordered another test using there difficult sequencing strategy. What an amazing transformation! From rubbish to near perfect and very easy to interpret sequences. Full credit to Macrogen – they did all of these retrials for free and in a small time frame.

The lesson is: If you have disappointing sequence results, don’t become despondent too quickly as there maybe a solution.