Sample preparation for sequencing - (Feb/16/2007 )
Hi all...
Someone out there can tell me how we send the samples for sequencin??? i know if it is a cloned product we can send it in the form of purified plasmid, wat i want to know is when we air dry the purified sample we have to be careful so tell me how many minutes or the time period i have to air dry the sample as the lyophilizer in our lab is not working properly so i can only do air dry....someone can help me out....
Thanks a lot....
Do you have to dry your samples? i normaly send it in water which has the concentration required by the company. It takes 2 days and works quite well.
Someone out there can tell me how we send the samples for sequencin??? i know if it is a cloned product we can send it in the form of purified plasmid, wat i want to know is when we air dry the purified sample we have to be careful so tell me how many minutes or the time period i have to air dry the sample as the lyophilizer in our lab is not working properly so i can only do air dry....someone can help me out....
Thanks a lot....
luckily we do sequencing by our own. after PCR and purification we dried the DNA sample for 15 min.
if you want to sent your plasmid DNA sample for sequencing ............i think normal procedure will be ok that is 5 to 10 min air dry on desk.
Someone out there can tell me how we send the samples for sequencin??? i know if it is a cloned product we can send it in the form of purified plasmid, wat i want to know is when we air dry the purified sample we have to be careful so tell me how many minutes or the time period i have to air dry the sample as the lyophilizer in our lab is not working properly so i can only do air dry....someone can help me out....
Thanks a lot....
Normally it depends on the company if they need lyophilized samples or samples in water/TE-buffer. Also the concentration needed in the latter is indicated, which you have to follow relative exactly (too much is as bad as too low concentration). We leave our samples often over night at room temperature in a safe place (our speedvac I do not trust anymore but this is a special problem).
So look at the intructions, they are often detailed and sufficient for all problems.
We used to leave the samples after precipitation in the 92C heat block for a min. for it to completely dry out. Then resuspend in the desired buffer and once again place it in hot block 92C for 2 min. then load into sequencer.