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

2. Report

3. 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

4. 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:
©2002 by American Society of Plant Biologists

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

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

7. Automated dideoxy DNA sequencing with fluoresncent ddNTPs

8. Next generation sequencing: Illumina/Solexa sequencing

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

10. 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.