Run restriction digestion: TA's will take the pictures for you. Review some points Sequencing techniques Analyze sequencing data

Report

Absorbance (4μl in 400μl water) = 0.05 Calculating the volume of miniprep plasmid DNA solution for your restriction assay Absorbance (4μl in 400μl water) = 0.05 Concentration = 0.05 * 50 (μg/ml) * 100= 250 μg/ml =250 ng/μl c1*v1 = c2 * v2 1's = stock solution c1= 250 ng/μl v1= ? c2 = 6 ng/μl (you needed 300ng in 50 μl= 6 ng/μl) v2 = 50 μl v1= 6*50/250 μl = 1.2 μl

Schematic of the map-based cloning process. Schematic of the map-based cloning process. Left, Typical 1-year mapping timeline for a mutation whose phenotype can be measured as the plants are growing. Right, Schematic of the five pairs of Arabidopsis chromosomes during critical stages of a sample mapping of a recessive mutation on chromosome 5 in the Col-0 background. Jander G et al. Plant Physiol. 2002;129:440-450 ©2002 by American Society of Plant Biologists

TAIR worksheet: 8. Which are the genes on either side of GA20OX-1?

TAIR worksheet: 8. Which are the genes on either side of GA20OX-1? AT4G2510 TTA AT4G25420 CAT ATG AT4G25430 TGA

Automated dideoxy DNA sequencing with fluoresncent ddNTPs http://www.mun.ca/biology/scarr/Fluorescent_didoxy_sequencing.html http://www.dnassequencing.com/2011/01/07/automated-dna-sequencing-6/

Next generation sequencing: Illumina/Solexa sequencing

Next generation sequencing: Illumina/Solexa sequencing Advantages: Fast Cheap (overall) Can be used for RNA sequencing and quantification Disadvantages: Sequence reads are short

BLAST The Basic Local Alignment Search Tool (BLAST) finds regions of local similarity between sequences. The program compares nucleotide or protein sequences to sequence databases and calculates the statistical significance of matches. BLAST can be used to infer functional and evolutionary relationships between sequences as well as help identify members of gene families. http://blast.ncbi.nlm.nih.gov/blast.cgi