Practical notes

Question 1

Beer-Lambert Law

Answer 1

A=ϵcl or A=log (Io/I), i.e., log (light intensity out/ in)
(ϵ in litres per Mol per cm)

Question 2

Lower pKa means…

Answer 2

Lower pKaH-> stabler anion-> better leaving group
1A= 0.1nm

Question 3

Using databases

Answer 3

If multiple proteins match a query sequence 100%, you can BLAST them and select matches that are related to the query protein. After that the longest natural (some are marked as engineered and likely to be of less use/relevance) sequence will probably be most useful.
cDNA is more useful when working with protein sequences than genomic as it excludes any alternative splicing possibilities/introns etc, hence guaranteeing the correct isoform for your query protein.
cDNA is translated in 6 frames (3 reading frames in both directions) and the longest continuous reading frame without stops/nonsense selected as the sequence could have been entered flipped/ don’t know reading frame from the start. After the correct reading frame is found, the sequence before the start codon can be discounted. The sequence can be double checked by comparing it with sequences in existing papers, running PCRs with random primers in the middle (discount possibility of polymerase falling off and getting truncated seq).