Today House Keeping Plasmid extraction, EcoRI digest PCR plants 15.ppt Bianca/microarrays (gels)

REPORTS

PARASITES AND SNAIL BIOLOGY “identity, possibilities” phylogenetics “intentions” transcriptomics PCR rDNA/mito Bioanalyzer DNA-free, direct sequencing gel electrophoresis nanodrop spec Sequence ID (BLAST) editing Phylogenetics electrophoresis RT-PCR gel CTAB Trizol TA cloning, B/W screening M13 sequencing Primer design, walking Qiagen plasmid extraction Restriction digests DNA RNA GenBank submission + Sevilleta snails

Snails: COI: complete sequences for S1 and S4 16S: complete sequence for S1 (partial S4) Failed reactions (possible reasons?) Generated amplicons from SS1 And SS2 another round of direct sequencing. Parasite: clones: Plasmids with inserts? Extract AND digest

LABEL YOUR PLASMID SAMPLES: initials and (e.g CA-1; CA-2; CA-3) INSERT CHECK BY RESTRICTION DIGEST SET UP restriction digestion for insert check: Label your tubes, initials and (e.g CA-1; CA-2; CA-3) Combine in a master mix: 4.5 microliter of EcoRI (storage buffers of restriction enzymes can inhibit reaction if >10% of final reaction volume) 4.5 microliter of 10x buffer 30 microliter of milliQ water, mix, aliquot 13 microliter to 3 tubes each. Add 2 microliter of plasmid sample Incubate >30 minutes, 37C. Run 1% agarose gel Inspect for presence absence of plasmids, inserts. LABEL YOUR PLASMID SAMPLES: initials and (e.g CA-1; CA-2; CA-3)

Plant PCR

15min.ppt