Background Info:
http://dwb4.unl.edu/Chem/CHEM869N/CHEM869NLinks/www.dur.ac.uk/~dbl0www/Staff/Croy/cDNAfigs.htm
mix:
water
reaction mix
RNA
enzyme (keep on ice)
add RNA
tap it
PCR
Thursday, May 27, 2010
Protein Extraction
http://www.piercenet.com/browse.cfm?fldID=FA97D803-6953-48E4-A7BD-6947D35FE83B
Basic Protocol:
Mix:
Lysis Buffer
Protease Inhibitor (comes in pellet --> vortex with buffer)
Phosphorase Inhibitor
add to pulverized tumor samples
homogenize while keeping tube on ice
spin down using vortex
Basic Protocol:
Mix:
Lysis Buffer
Protease Inhibitor (comes in pellet --> vortex with buffer)
Phosphorase Inhibitor
add to pulverized tumor samples
homogenize while keeping tube on ice
spin down using vortex
Wednesday, May 26, 2010
Tuesday, May 25, 2010
Monday, May 24, 2010
BLAST Algorithm and Substitution Matrix
http://en.wikipedia.org/wiki/BLAST#Algorithm
http://en.wikipedia.org/wiki/Substitution_matrix
http://en.wikipedia.org/wiki/Substitution_matrix
Sequence Alignment
http://en.wikipedia.org/wiki/Sequence_alignment
*know global vs local alignment, various programs: FASTA, genebank, dot matrix-hits on the main diagonal determines sequence similarity
*know global vs local alignment, various programs: FASTA, genebank, dot matrix-hits on the main diagonal determines sequence similarity
Poisson Distribution
http://en.wikipedia.org/wiki/Poisson_distribution
http://bioinfo.mbb.yale.edu/course/classes/c4/c4-p1.html
"Having introduced the idea of approximating the binomial distribution with two distributions, each of which is applicable in a different regime of the value of p, lets consider the case where p is small (p0.1). First, let us perform the substitution l =np. The binomial distribution then becomes,
(7)
Now, consider the case where n grows to infinity and p shrinks to zero. Hopefully you appreciate the utility of the substitution that we made above, since we can force n to grow and p to shrink such that l =np remains constant. This is nice since nothing in the above expression will "blow up" for large n and/or small p. In this limit we get,
(8) for k=0,1,2,3,...
This expression is the Poisson distribution, and is useful in the situations where the probability of an occurrence is small and the number of "trials" (n) is large. For example, we might consider the probability of k adverse reactions to a test drug in a given sample of the population or the probability of registering k complaints about a particular product in a 1-hour period or the probability of finding k point mutations in a given stretch of nucleotides. Though the Poisson distribution is essential to application and you will doubtless see it again, we will leave it now to discuss the other binomial-approximating continuous distribution. "
http://bioinfo.mbb.yale.edu/course/classes/c4/c4-p1.html
"Having introduced the idea of approximating the binomial distribution with two distributions, each of which is applicable in a different regime of the value of p, lets consider the case where p is small (p0.1). First, let us perform the substitution l =np. The binomial distribution then becomes,
(7)
Now, consider the case where n grows to infinity and p shrinks to zero. Hopefully you appreciate the utility of the substitution that we made above, since we can force n to grow and p to shrink such that l =np remains constant. This is nice since nothing in the above expression will "blow up" for large n and/or small p. In this limit we get,
(8) for k=0,1,2,3,...
This expression is the Poisson distribution, and is useful in the situations where the probability of an occurrence is small and the number of "trials" (n) is large. For example, we might consider the probability of k adverse reactions to a test drug in a given sample of the population or the probability of registering k complaints about a particular product in a 1-hour period or the probability of finding k point mutations in a given stretch of nucleotides. Though the Poisson distribution is essential to application and you will doubtless see it again, we will leave it now to discuss the other binomial-approximating continuous distribution. "
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