progress report 2

29/7/2010

After we analyzed the sequencing results, actually there were some problems.

KAS I: - we manage to get the correct insert (KAS I) in pQE30. however, when we analyze the nucleotides sequence, one of the nucleotide was missing at the primer sequence. so, we already sent the same plasmid for re-sequencing for confirmation. there are 2 probabilities; first, problems maybe occured during the sequencing process, or problems with the KAS I primer which the BioNeer did not give the complete primer sequence.

KAS II: - basically we manage to get the correct insert (KAS II) into pQE30. The problems also the same as KAS I, but, instead of missing one nucleotide, it happen to be an extra nucleotide at the primer. so, for confirmation, we decided to send for re-sequencing again and compare with other clones.

KAS III : - we not able to sequence this pQE30+KAS III because we got the wrong clone. maybe KAS III was inside the pGEMT vector. thus, we planned to digest the pGEMT-KAS III and ligate it with pQE30 into E.coli JM109 again and send for sequencing.

That's all.

More progress will be updated soon.

-HADI /HANI-