Are nucleic acids (DNAs and RNAs) positively charged, negatively charged, or neu

Question

Are nucleic acids (DNAs and RNAs) positively charged, negatively charged, or neutral in a basic buffer such as TAE? What is the direction of migration for a DNA or RNA molecule in an electric field? What kind of gel (starch, polyacrylamide, or agarose) will be used for DNA electrophoresis? Why is agarose gel more appropriate than polyacrylamide gel for DNA analysis? Agarose gel has bigger pore size which is more suitable for big DNA molecules. Assuming the PCR and Smal digestion reactions you did in last week's lab all work, what would you see in the agarose gel you will run: Lane sample (5 1) (5011) (50 il) (50 ul) 100 bp DNA marker 2PCR product by group member 1 PCR product by group member 2 PCR product by group member 3 MacBook A

Explanation / Answer

1) TAE is a basic buffer that has Tris base, Acetic acid and EDTA. The primary functions of this buffer is to,

Hence the nucleic acids are negatively charged in the basic buffers such as TAE buffer.

2) The DNA & RNA molecule are negatively charged molecule due to the phosphate backbone. Hence these molecules will move towards the positively charged anode. So the side of gel which has wells is placed towards the cathode and then the molecules migrate towards the anode

3) Agarose gel is the most suited gel for the separation of the nucleic acids.

4) The size of DNA and RNA is larger as compared to the proteins and agarose gels have bigger pore size and hence these are most suited for the nucleic acid separation.

5) As the information about the restriction sites that are present in the DNA which needs to be digested. So, I am providing you the basic information.

PCR is the polymerase chain reaction which amplifies the selected fragment using the specific primers.

The digestion with the Sma1 will cut the PCR product into fragment based on how many sites are present in the PCR product. So let’s assume there is one site present, the enzyme will work there and cleave the product into two fragments. Now if the mass of both fragment coincide then one band will be seen and if different, two will be seen on the gel. Any other band should be considered due to the non-specific amplification during PCR.

LANES

VISUALIZATION ON THE GEL

1.

100 bp ladder will resolves in 10bands ranging from 100 bp to 1000 bp.

2.

2 bands

3.

2 bands

4.

2 bands

Provide more information for detailed answer.

LANES

VISUALIZATION ON THE GEL

1.

100 bp ladder will resolves in 10bands ranging from 100 bp to 1000 bp.

2.

2 bands

3.

2 bands

4.

2 bands

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