| Features of stem cells | - unspecialised
- develop into other specialised cells
-divide renew themselves |
| Multipotent stem cells | This type of cells can specialise into few different types of body cells
-adults mammals - bone marrow |
| Unipotent stem cells | Can only specialise into one type of body cell
-found in adult mammals |
| Induced pluripotent stem cells | a unipotent cell is forced/induced to become a pluriportent cell |
| Ways to remeber all your stem cells | TOTally Pretty MULTIcoloured Unicorn
totipoten /all Pluripotent/all except placenta Multipotent/few Unipotent/1 |
| What is differentiation | specilaisation of stem cells
switching genes on |
| What is a promoter region | upstream of a gene |
| 3 ways to create DNA fragments(isolating genes) | 1. cDNA
2. Restriction enzyme
3. Gene machine |
| What is the process of making CDNA | Reverse transcriptase is added to mrna to make a template
free DNA nucleotided mkae complementary base pairs with mRNA , reverse transcriptase joins the backbone. Copies of the CNA bound are copied cDNA strands join by DNA polymerase |
| The cut / reverse transcriptase | A gene is cut at the restriction site by a restriction enzyme
. The enzymes cuts at specific sequence of bases known as a restriction site , this is palindromic sequence, the enzyme breaks the phosphodiester bonds leaving sticky ends |
| Example of DNA amplification / in vitro cloning | Polymerase chain reaction |
| What are key temperatures important for PCR | 90-50-70 |
| What is PCR | At 90 degrees DNA is heated to break hydrogen bonds , it then cooled at 50 degrees primers are added to the ends of each single strands, it is then heated to 70 degrees free nucleotides are added so complemantry base pairs ,DNA polymersae joins back bones so two DNA strands produced |
| In Vivo cloning is what ? | cut cut stick |
| Cut cut stick theory | Cut out gene - by restriction enzyme theory leaving sticky ends
Cut out gene at bacteria - restriction enzyme theory of plasmid
Stick - sticky ends join together DNA ligase forms back bone creating recombinant DNA |
| How do you insert into bacteria and grow and find it | HEAT SHOCK AND ICE COLD SHOCK with CaCL2 so bacteria takes up plasmid
Find by marker gene and grow on petri dish |
