The separation of chapter 1 quality grain DNA, purely turn and authenticate
The first upright conduct say
Pass the reorganization DNA technique to an useful purpose DNA part, send into be subjected to body cell in carry on breed with expression of tool call carry a body(Vector).The germ quality grain reorganizes to in common usely carry a body in the DNA technique.
Quality grain(Plasmid) the stable gene which is the outside of a kind of chromosome, size from 1-200 kbs not etc., for double chain, shut the DNA member of wreath, and exist by super and spiral appearance in host cell.The quality grain is main to discover at the germ and put line germ and fungi cell in, it has an independent replication and transcribes ability, can in the son generation the cell keep Heng settle of copy a number, and express the heredity information take.The replication of quality grain with transcribe and depend on some Maos and protein which code at the host cell, if leave host cell, can’t survive, and the host even have no them and can also normally survive.The existence of the quality grain makes the host have some additional characteristics, such as to anti- sex of the antibiotics etc..F quality grain(call F factor or property grain again), R quality grain(drug-resistant factor) with Col quality grain’s(produce the E. coli plain Continue reading »
Chapter 2 DNA Mao slice and gel electricity Yong
The first upright conduct say
A. Slice Mao Mao to slice analysis inside DNA restriction
Limitting to slice Mao inside sex can especially foreign land’s combine be called restriction Mao to identify the DNA sequence of sequence at 1 inside or its neighborhood of especially the different order up, and incise a pair of chains DNA.It can is divided into 3 types: Ⅰ and Ⅲ Mao are in same protein member and have already incise with polish(A Ji turn) a function and depend on at the ATP existence.The Ⅰ Mao combine in identify a DNA that point combines to random incise to identify a point not distance, but the Ⅲ Mao incise DNA member in identifying a point, then the solution leave from the bottom thing.Ⅱ Constitutes to°from two kinds of Maos: In order to limit to slice nucleic acid Mao(restriction Mao) inside sex 1 kind, it incise some an especially different pit Mao sour sequence; Another turn Mao for the independent A Ji, it polish same identify a sequence.Sliced Mao to get in the member gram the Long inside the restriction in Ⅱ extensively applied, they are the foundations which reorganizes DNA.The great majority Ⅱ restriction Mao identifies length to check Mao sour reply for 4-6 symmetry especially different pit Mao sour sequence(such Continue reading »
Chapter 3 E. coli feels the making and conversion of Tai cell
The first upright conduct say
Under the natural condition, a lot of quality grains all can pass a germ coalescence a function to transfer a new host, but generally lack in the artificial set up of the quality grain carry the body this kind transfer the essential mob gene, so can’t complete by oneself to transfer from a cell to another coalescence of cell.If need to carry a quality grain a body to transfer into be subjected to a body germ, need inducement to be subjected to a body germ to produce a kind of brief feeling Tai to take outside source DNA.
The conversion(Transformation) leads the outside source DNA member in to be subjected to body cell and make it acquire a kind of means of new hereditary form, it is a microorganism heredity, member heredity, genetic engineering etc. research realm of basic experiment technique.
Convert what process use is generally been the variation stub that the restriction polishes system blemish by body cell, then don’t contain to slice the mutation body(R ˉ , M ˉ ) that Mao and A Ji turn Mao inside restriction, it can is tolerant of outside source DNA member to get into a body combine stability ground the heredity is to the posterity.Be subjected to body cell through some processings of special methods(like the electric shock method, CaCl2, the RbCl(KCl) equalizes to learn to try a method), the change of palliation Continue reading »
Chapter 4 RNA withdraws to synthesize with cDNA
The first upright conduct say
Withdraw mRNA from truely the pit the living creature of organization or cell, converse to record to synthesize cDNA through the reaction of Mao Cu the first chain and the second chain, double chain cDNA with carry a body conjunction, then convert to expand, can immediately acquire a cDNA library, set up of the cDNA library can used for true pit living creature gene of structure, expression with adjust the analysis for control;Compare the cDNA can make sure that containing sub- existence and understanding processes to wait a series of problem after transcribe with corresponding a gene set a DNA sequence difference.In fine the cDNA synthesize to have become to really check the basic means of molecular biology nowadays with gram Long.From 70’s middle period head the example cDNA gram the Long publish, have already developped many methods that the exaltation cDNAs synthesize an efficiency, and consumedly improved to carry system to govern, cDNA’s synthesizing to try have already commercialize currently.The cDNA synthesize and gram the basic step of the Long include with the reversal record Mao to synthesize cDNA the Continue reading »
The conjunction of chapter 5 reorganization quality grain, conversion and sieving
The first upright conduct say
The quality grain has stable credibility and operates a simple advantage.If want a gram of DNA part(<10 kbs) with smaller Long and structure simple, the quality grain wants to compare other any carry bodies to all be good friends with.Carry a body in the quality grain up carry on gram Long, say from the principle that is easy, use first to slice Mao to incise a quality grain DNA and purpose DNA part inside restriction, then the body outside make both connect with each other, use again to gain reorganization quality grain to convert a germ and then complete.But in actually the work, how classify to insert a reorganization quality grain of have the outside source DNA with insert but the oneself wreath turn of carry body member is more difficult.Pass to adjust a conjunction to respond Chinese and Foreign source DNA part and carry the density comparison of body DNA, can carry the oneself wreath of body to turn restriction at certain degree under, can also adopt some special gram Long strategy further, if carry a body to turn to the phosphoric acid etc. come utmost lower oneself wreath of carry the body to turn, can also make use of genetics means,such as Continue reading »
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