run restriction digestion: ta's will take the pictures for you. review some points sequencing...

Run restriction digestion: TA's will take the pictures for you. Review some pointsSequencing techniquesAnalyze sequencing data

Report

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/μlv1= ?c2 = 6 ng/μl (you needed 300ng in 50 μl= 6 ng/μl)v2 = 50 μl

v1= 6*50/250 μl = 1.2 μl

Calculating the volume of miniprep plasmid DNA solution for your restriction assay

Schematic of the map-based cloning process.

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?

AT4G25420TTA

CAT

ATG

TGA

AT4G25430

AT4G2510

Automated dideoxy DNA sequencing with fluoresncent ddNTPs

http://www.mun.ca/biology/scarr/Fluorescent_didoxy_sequencing.htmlhttp://www.dnassequencing.com/2011/01/07/automated-dna-sequencing-6/

Next generation sequencing: Illumina/Solexa sequencing

Next generation sequencing: Illumina/Solexa sequencing

Advantages: FastCheap (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