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Definition
| Visualizes chagnes in the arrangement of genomic DNA in one cell type compared to another based on mobility on gel electrophoresis |
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Term
| How to compare rearranged vs unrearranged DNA |
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Definition
Clone DNA fragment containing rearrange gene
2) restriction map - to determine how it differes from unrearranged DNA
3)subclone - a smaller fragment containing site of recombination
4) Sequence - the rearranged DNA compare to sequence of unrearranged DNA, using bioinformatics likBLAST |
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Term
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Definition
Polymerase Chain Reaction
Amplifies rearranged DNA from populations of cells
2) gel electrophoresis can be used to determien the types of rearrangement that occcur in a different cell type |
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Definition
Can tell you the stages of immunological development
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Term
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Definition
| nonionic detergent dissolves the plasma membrane of the cells while leaving the nuclear membrane intact |
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Term
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Definition
| denatures proteins (such as DNases) and lyses nuclear membranes |
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Definition
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Definition
| use isopropanol, it's insoluble and precipitates |
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Definition
Tris-EDTA a weak buffer with EDTA that traps metals and thus inhibits enzymes like DNases
Used commonly when storing protein/dna to save them from being degraded by trace enzymes |
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Term
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Definition
pH extremes
enzymes
shear forces |
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Term
| There are 7 antigen receptor loci in lymphocytes, name them |
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Definition
T cells
alpha beta gamma sigma
B cells
kappa lambda |
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Term
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Definition
| catalyzes V-D-J rearrangement |
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Term
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Definition
recombination signal sequence
heptamer
spacer (12/23)
nonamer
Rearrangement occurs adajacent to heptamer sequence
V's flanked 3' with RSS
D's Flanked both sides
J's flanked 5' with RSS
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Term
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Definition
| only RSS with opposite spacer sizes can rearrange together |
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Term
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Definition
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Term
| How does VDJ recombination imprecise? |
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Definition
Exact site of recombination varies
TdT (terminal transferase)- can add a few random base pairs at the site of joining
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Term
| Light Chain joins which two segments? |
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Definition
V and J
Also, TdT does not add N regions to light chains rearranging segments |
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Term
| What causes the loss of nucleotides between rearranging segment DNA? |
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Definition
| Exonucleases cause for more diversity between V-D and D-J |
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Term
| What is a good 260/280 ratio? |
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Definition
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Term
| How to calculate concentration of a sample of DNA? |
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Definition
If ratio is >1.6
Then concentration is OD x 50ug/ml = concentration of DNA |
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Term
| One unit of restriction enzyme is? |
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Definition
| one unit of restriction enzyme is defined by the amount nccessary to cut 1 ug of plasmid DNA in one hour |
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Term
| What enables you to see DNA on a gel using UV light? |
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Definition
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Definition
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Term
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Definition
| total number of nonrepeating base-pairs inteh sample |
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Term
| Southern blot sensitivity |
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Definition
| 1x10^6 copies for one band |
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Definition
| catalyzes NBT and X-phosphate |
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Definition
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Definition
| substrate that turns blue |
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