Butterfly/moth-barcoding

Hi,

In the following weeks I have to do a barcoding for a certain moth-specimen, and I could amplify more 4-5 samples (I have less DNA-polymerase), if somebody would have sample. As you know, the barcoding is a molecular technique for identification of an animal specimen/individual. This is a 3steps-procedure with DNA-extraction, PCR and sequencing. During the PCR, we ampify the gene encoding the mitochondrial cytochrome-oxidase subunit I (to be more precisely a part of it) and this DNA-fragment will be sequenced. I have primers (oligonucleotides) only for Lepidoptera-species.

So, if somebody has some fresh (maximum 1-2 years old) but problematic, unidentifeid specimen (or one member of a problematic pair, let's say Leptidea sinapis/reali), I could help her/him. What kind of tissue do I have to send, if you are interested in this? So, generally the legs are used for this aim, but at micromoths the best is the abdomen, because the low mass of legs. If you want to take away the abdomen for genitalic study (so, you want to do a microscopic slide from the genital structures), then I propose, cut please the proximal segments of the abdomen, and it is worth to send these tissue-pieces to me.

We can get ca. 620-640 basepair length from the standard 658 basepair length with 1 sequencing. If you would like to get the full-length 658 bp sequence, that is plus one sequencing, but after my opinion, to an identification the 620-640 is enough (since that is ca. 96% of the 658-size, if we calculate with 630)

If you are interested in this, please write to me in PM/to my mail-address (also in German ), especially important to talk about the storage condition, the "history" of the given specimen. The age is also very important. We cannot get barcode sequence from too old specimen, and at my present conditions it is impossible.

Thank you
Kornél

PS: Just some pics, one about the agarose gel, in which I run the amplified mtCOI amplicon. You can see 5 lanes, the first from left is the DNA-ladder, or DNA-marker, the next three lane is a tryings or probes with different specimens collected in 1984.. They were negative, as you can see. And the last lane shows a positive control, a freshly collected Colias croceus. It is ok.

Next picture is a part of a chromatogram that is made at the sequencing.

If you will get the sequence from me, here you can perform the identification (simple copy/paste) and next surface will give out list after similarity
BOLD Systems: Management & Analysis - Identification Request