Lecture 23-24

Question 1

restriction enzymes

Answer 1

• bacterial immune system
• pallidromic sequences
• 􏰄 􏰅􏰪􏰓􏰔􏰗􏰨􏰗􏰔􏰇􏰮􏰯􏰰􏰇􏰓􏰖􏰤􏰎􏰖􏰙􏰔􏰢􏰓􏰌􏰚􏰓􏰚􏰇􏰨􏰎􏰏􏰇􏰔􏰙􏰐􏰐􏰗􏰖􏰦􏰇􏰮􏰯􏰰􏰇􏰐􏰎􏰇􏰚􏰐􏰙􏰤􏰑􏰇􏰳􏰇􏰔􏰢􏰎􏰖􏰄 􏰅􏰪􏰓􏰔􏰗􏰨􏰗􏰔􏰇􏰮􏰯􏰰􏰇􏰓􏰖􏰤􏰎􏰖􏰙􏰔􏰢􏰓􏰌􏰚􏰓􏰚􏰇􏰨􏰎􏰏􏰇􏰔􏰙􏰐􏰐􏰗􏰖􏰦􏰇􏰮􏰯􏰰􏰇􏰐􏰎􏰇􏰚􏰐􏰙􏰤􏰑􏰇􏰳􏰇􏰔􏰢􏰎􏰖􏰓

Question 2

restrction mapping

Answer 2

􏰰􏰇􏰚􏰓􏰏􏰗􏰓􏰚􏰇􏰎􏰨􏰇􏰚􏰗􏰖􏰦􏰢􏰓􏰇􏰳􏰇􏰤􏰎􏰙􏰍􏰢􏰓􏰇􏰤􏰗􏰦􏰓􏰚􏰐􏰚􏰇􏰐􏰎􏰇􏰙􏰖􏰤􏰓􏰏􏰚􏰐􏰌􏰖􏰤􏰇 􏰐􏰕􏰓􏰇􏰢􏰎􏰔􏰌􏰐􏰗􏰎􏰖􏰚􏰇􏰎􏰨􏰇􏰏􏰓􏰚􏰐􏰏􏰗􏰔􏰐􏰗􏰎􏰖􏰇􏰚􏰗􏰐􏰓􏰚􏰇􏰏􏰓􏰢􏰌􏰐􏰗􏰟􏰓􏰇􏰐􏰎􏰇􏰓􏰌􏰔􏰕􏰇 􏰎􏰐􏰕􏰓􏰏􏰇􏰎􏰖􏰇􏰌􏰇􏰮􏰯􏰰􏰇􏰨􏰏􏰌􏰦a series of single and double digests to understand the locations of restriction sites relative to each other on a DNA strand

Question 3

Strategies to making a map

Answer 3

-determine if it is linear or circular
-make single digest maps and then combine them
-keep track of all sizes/ distances
􏰄 􏰆􏰎􏰣􏰪􏰌􏰏􏰓􏰇􏰚􏰗􏰖􏰦􏰢􏰓􏰇􏰌􏰖􏰤􏰇􏰤􏰎􏰙􏰍􏰢􏰓􏰇􏰤􏰗􏰦􏰓􏰚􏰐􏰇􏰚􏰗􏰲􏰓􏰚􏰇􏰐􏰎􏰇􏰤􏰓􏰐􏰓􏰏􏰣􏰗􏰖􏰓􏰇 􏰐􏰕􏰓􏰇􏰗􏰤􏰓􏰖􏰐􏰗􏰐􏰑􏰇􏰎􏰨􏰇􏰐􏰕􏰓􏰇􏰍􏰌􏰖􏰤􏰚
􏰄 􏰹􏰓􏰓􏰪􏰇􏰐􏰏􏰌􏰔􏰵􏰇􏰎􏰨􏰇􏰌􏰢􏰢􏰇􏰚􏰗
􏰄 􏰄􏰌􏰵􏰓􏰇􏰚􏰗􏰖􏰦􏰢􏰓􏰇􏰤􏰗􏰦􏰓􏰚􏰐􏰇􏰣􏰌􏰪􏰚􏰇􏰐􏰕􏰓􏰖􏰇􏰔􏰎􏰣􏰍􏰗􏰖􏰓􏰇􏰐􏰕􏰓􏰣
􏰄 􏰮􏰗􏰦􏰓􏰚􏰐􏰚􏰇􏰐􏰕􏰌􏰐􏰇􏰦􏰗􏰟􏰓􏰇􏰨􏰓􏰠􏰓􏰏􏰇􏰍􏰌􏰖􏰤􏰚􏰇􏰌􏰏􏰓􏰇􏰓􏰌􏰚􏰗􏰓􏰏􏰇􏰐􏰎􏰇􏰣􏰌􏰪􏰇􏰨􏰗􏰏􏰚􏰐
􏰄 􏰆􏰎􏰣􏰪􏰌􏰏􏰓􏰇􏰚􏰗􏰖􏰦􏰢􏰓􏰇􏰌􏰖􏰤􏰇􏰤􏰎􏰙􏰍􏰢􏰓􏰇􏰤􏰗􏰦􏰓􏰚􏰐􏰇􏰚􏰗􏰲􏰓􏰚􏰇􏰐􏰎􏰇􏰤􏰓􏰐􏰓􏰏􏰣􏰗􏰖􏰓􏰇 􏰐􏰕􏰓􏰇􏰗􏰤􏰓􏰖􏰐􏰗􏰐􏰑􏰇􏰎􏰨􏰇􏰐􏰕􏰓􏰇􏰍􏰌􏰖􏰤􏰚
􏰄 􏰹􏰓􏰓􏰪􏰇􏰐􏰏􏰌􏰔􏰵􏰇􏰎􏰨􏰇􏰌􏰢􏰢􏰇􏰚􏰗􏰲􏰓􏰚􏰱􏰤􏰗􏰚􏰐􏰌􏰖􏰔􏰓􏰚

Question 4

Agrose gel electrophoresis

Answer 4

common for long DNA fragments 􏰄 􏰅􏰐􏰌􏰖􏰧􏰌􏰏􏰧􏰇􏰤􏰓􏰥􏰚􏰇􏰨􏰎􏰏􏰇􏰍􏰌􏰖􏰧􏰚􏰇􏰙􏰦􏰇􏰐􏰎􏰇􏰩􏰈􏰊􏰝􏰊􏰊􏰊􏰇􏰍􏰦 􏰥􏰎􏰖􏰤􏰄 􏰅􏰐􏰌􏰖􏰧􏰌􏰏􏰧􏰇􏰤􏰓􏰥􏰚􏰇􏰨􏰎􏰏􏰇􏰍􏰌􏰖􏰧􏰚􏰇􏰙􏰦􏰇􏰐􏰎􏰇􏰩􏰈􏰊􏰝􏰊􏰊􏰊􏰇􏰍􏰦 􏰥􏰎􏰖􏰤􏰄 􏰅􏰐􏰌􏰖􏰧􏰌􏰏􏰧􏰇􏰤􏰓􏰥􏰚􏰇􏰨􏰎􏰏􏰇􏰍􏰌􏰖􏰧􏰚􏰇􏰙􏰦􏰇􏰐􏰎􏰇􏰩􏰈􏰊􏰝􏰊􏰊􏰊􏰇􏰍􏰦 􏰥􏰎􏰖􏰤
􏰄 􏰪􏰙􏰥􏰚􏰓􏰧􏰇􏰤􏰓􏰥􏰚􏰇􏰨􏰎􏰏􏰇􏰥􏰌􏰏􏰤􏰓􏰏􏰇􏰨􏰏􏰌􏰤􏰫􏰓􏰖􏰐􏰚􏰝􏰇􏰔􏰌􏰖􏰇􏰍􏰓􏰇􏰙􏰚􏰓􏰧􏰇􏰐􏰎􏰇 􏰨􏰗􏰖􏰤􏰓􏰏􏰦􏰏􏰗􏰖􏰐􏰇􏰍􏰌􏰔􏰐􏰓􏰏􏰗􏰌􏰥􏰇􏰚􏰐􏰏􏰌􏰗􏰖􏰚􏰄 􏰪􏰙􏰥􏰚􏰓􏰧􏰇􏰤􏰓􏰥􏰚􏰇􏰨􏰎􏰏􏰇􏰥􏰌􏰏􏰤􏰓􏰏􏰇􏰨􏰏􏰌􏰤􏰫􏰓􏰖􏰐􏰚􏰝􏰇􏰔􏰌􏰖􏰇􏰍􏰓􏰇􏰙􏰚􏰓􏰧􏰇􏰐􏰎􏰇 􏰨􏰗􏰖􏰤􏰓􏰏􏰦􏰏􏰗􏰖􏰐􏰇􏰍􏰌􏰔􏰐􏰓􏰏􏰗􏰌􏰥􏰇􏰚􏰐􏰏􏰌􏰗􏰖􏰚􏰄 􏰪􏰙􏰥􏰚􏰓􏰧􏰇􏰤􏰓􏰥􏰚􏰇􏰨􏰎􏰏􏰇􏰥􏰌􏰏􏰤􏰓􏰏􏰇􏰨􏰏􏰌􏰤􏰫􏰓􏰖􏰐􏰚􏰝􏰇􏰔􏰌􏰖􏰇􏰍􏰓􏰇􏰙􏰚􏰓􏰧􏰇􏰐􏰎􏰇 􏰨􏰗􏰖􏰤􏰓􏰏􏰦􏰏􏰗􏰖􏰐􏰇􏰍􏰌􏰔􏰐􏰓􏰏􏰗􏰌􏰥􏰇􏰚􏰐􏰏􏰌􏰗􏰖􏰚

Question 5

Basic component of common PCR method

Answer 5

• template DNA
• oglinucleotide primers
• dNTP
• Taq plymerase
• suitable buffer solution

Question 6

number or PCR copies=

Answer 6

􏰻􏰈􏰖 􏰷 􏰈(2^N-2N)X

Question 7

TAQ=

Answer 7

-HALF LIFE IS 40 MINUTES
-misincorporated nucleotides are not removed
adds A to 3’ end

Question 8

PFU

Answer 8

longer half life

• misincorporated nucleotides are removed
• no A addition