4 March 2014 DNA precipitation According to picture of the last PCR, the gene M and S have base pair

500bp approximately, it means 75 ng, btw the band seem like 2-3x time more strong, it means 150-225 ng,, for the 25l volume,, so we guess if we use 50 ml it could be around 500ng Step 1* 500ng,,,of DNA contain on PCR tube, @50 l. It mean 0,5 l 0,5 unit, because 1 microgram DNA is cut by RE(restriction enzyme) 1 unit/1 hour. Questions : If N1a III 10.000 U/ml = 10 u/l = How much microliters we using for 0.5 unit ???? Answer * 0.05 l,, But it is difficult to get this amount with micropippete, so we use 0.05 microliter in the real jikken.

March 2014 DNA precipitation..(for 50 l pcr tube)

First prepare eppendorf tube and take with micropipete 50 l Genomic DNA from PCR tube before, and then perform this : -3M sodium Acetate 1.65 l -Ethachinmate 0.5 l -add ethanol 2x time of volume 100 l -vortex 2-3 times -12K x g 5 min (12.000 speed of sentrifuge) -Suction the etanol liquid,and save the pellet in the bottom of effendorf tube. -Dry the pellet in the air.within 1 hours -Further more doing,,next procedure --- N1a digestion 2. N1a III digestion step 1 * NE-B4 BSA N1a III DNAadd destilled water Total 3 l 0,3 l .. l .. l 30 l

Incubate @ 37 celsius cold for 2 hours, wait Load the sample into 3 % agarose XP nippon gene (0.03 x 25 ml of TAE=0,75 gr of powder), Step2* agarose gel , Make loading 6x buffer DNA genomic GL ladder 100 Step 3*Electrophoresis Step 4*Staining with etylen bromide 3% composition : 2 l : 10 l : 5 l

Comparison chart</> Embed this chart

DNA
Stands for DeoxyriboNucleicAcid

RNA

RiboNucleicAcid A single-stranded chain of alternating A nucleic acid that contains the genetic phosphate and ribose units with the bases instructions used in the development and Adenine, Guanine, Cytosine, and Uracil functioning of all modern living organisms Definition bonded to the ribose. RNA molecules are (scientists believe that RNA may have been involved in protein synthesis and the main genetic material in primitive life sometimes in the transmission of genetic forms). information. Transfer the genetic code needed for the Medium of long-term storage and Function creation of proteins from the nucleus to the transmission of genetic information ribosome. The helix geometry of DNA is of B-Form. The helix geometry of RNA is of A-Form. Unique DNA is protected in the nucleus, as it is RNA strands are continually made, broken Features tightly packed. DNA can be damaged by down and reused. RNA is more resistant to exposure to ultra-violet rays. damage by Ultra-violet rays. A single-stranded molecule in most of its Predominant Double- stranded molecule with a long chain biological roles and has a shorter chain of Structure of nucleotides nucleotides Deoxyribose sugar; phosphate backbone; Ribose sugar; phosphate backbone. Four Bases & Four bases: adenine, guanine, cytosine and bases: adenine, guanine, cytosine, and Sugars thymine uracil Pairing of A-T(Adenine-Thymine), G-C(GuanineA-U(Adenine-Uracil), G-C(GuanineBases Cytosine) Cytosine) Deoxyribose sugar in DNA is less reactive Ribose sugar is more reactive because of because of C-H bonds. Stable in alkaline C-OH (hydroxyl) bonds. Not stable in Stability conditions. DNA has smaller grooves, which alkaline conditions. RNA has larger makes it harder for enzymes to "attack" grooves, which makes it easier to be DNA. attacked by enzymes. RNA is synthesized from DNA when Propagation DNA is self-replicating. needed.

Summary of Differences Between DNA and RNA

1. 2. 3. 4. 5. DNA contains the sugar deoxyribose, while RNA contains the sugar ribose. The only difference between ribose and deoxyribose is that ribose has one more -OH group than deoxyribose, which has -H attached to the second (2') carbon in the ring. DNA is a double stranded molecule while RNA is a single stranded molecule. DNA is stable under alkaline conditions while RNA is not stable. DNA and RNA perform different functions in humans. DNA is responsible for storing and transferring genetic information while RNA directly codes for amino acids and as acts as a messenger between DNA and ribosomes to make proteins. DNA and RNA base pairing is slightly different, since DNA uses the bases adenine, thymine, cytosine, and guanine; RNA uses adenine, uracil, cytosine, and guanine. Uracil differs from thymine in that it lacks a methyl group on its ring.