Pineapple DNA Extraction - Very low or no yield (Apr/15/2007 )
Hi, I have been trying to extract the genomic DNA of pineapple but failed so far, as either the DNA yield is very low (despite resuspending my DNA in a much lesser volumn of dH2O than that suggested by the kit I'm using) or there is none at all. I'm using Promega's DNA Extraction kit by the way, following the protocol of plant DNA extraction.
What I've tried so far includes adding SDS together with the kit's nuclei lysis buffer (the initial cell wall lysing step) and proteinase K (to breakdown any enzyme which might degrade the DNA or inhibit PCR in later steps) after RNAse treatment.
I'm planning on getting the Qiagen's DNA extraction kit specifically for plants.
Any experience of how to extract DNA from pineapple, as well as the efficiency of the Qiagen kit?
kits usually give low yield, so if you want a large amount of DNA it is better to go for CTAB macro extraction.
Qiagen kits usually give good results, how much DNA are you expecting and how much are you getting?
it's clear that pineapple needs extra concentration to extract its DNA ot try to use more tissue volume (more pineapple) with the same kit
What material are you starting with? The fruit is probably a bad choice. New leaves would be my first choice.
So true, the fruit is horribly acidic. And that will throw off the pH of the buffers, which inturn will affect the DNA extraction kit, the kit binds DNA in a pH and salt concentration dependent manner
I always have this thinking whereby for plants/fruits, use CTABS method for best DNA extraction. Is it really true? I mean... i know CTABs is widely used for DNA extraction from plants. SO perhaps you can try to use CTAB method.