RFLP is restriction fragment length polymorphism. This is a method of exploiting the diferences in sequence of nucleotides between individuals. This can be done on any DNA, from what ever animal, plant or bacteria you choose. It can even be done on viruses. Restriction enzymes are endonucleases (nucleotide enzymes that recognize a specific sequence and cleave the DNA at that site). If you have 2 different DNA strands, one of the quickest tests that can be done to recognize the differences between them is to subject them to RFLP. Because the sequences are diffferent, the restriction enzymes will cut them up in different ways: Maybe the first one will be cut into 10 segments of different lengths whereas the second will be cut into 50 segments of smaller size. Then when you run these fragments on a gel, you will see that they appear different.

TRFLP is a similar method used to examine the DNA of bacteria to determine differences (possibly mutations). TRFLP requires PCR amplification (Polymerase chain reaction). You can use these techniques to identify SNPs (single nucelotide polymorphisms).

Equipment necessary includes the reagents, restriction enzymes, the DNA samples and other test samples of different lengths for comparison. SDS-PAGE which stands for polyAcrylamide Gel Electrophoresis.

These techniques would tell you some subtle differences between the nucleotide sequence of 2 or more different samples of DNA. The restriction enzymes will break each sample of DNA into different number and length of fragments because of the changes in the sequences between the samples. The restriction enzymes usually recognize specific palindromic sequences on the DNA. Once the samples are mixed with the restriction fragments and digested, the samples are placed in the wells of the gel and current is passed through the gel to make the different fragments migrate down the gel according to size. Then you can read from the gel how many and what size fragments were created by the enzymes

RFLP can be done directly on genomic DNA or on PCR products however T-RFLP is specifically designed to be used on PCR amplicons of conserved region genes. T-RFLP can help in enumeration of endophytes based on variable number of fragments generated hence T-RFLP is advisable however it will require one or two labelled primers and Capillary electrophoresis. RFLP can be done with only PCR, Agarose gel.

Dear Pooja Singh

Pls. find the attached files and the references,

Hoping this will be helpful to your project, good luck

Regards

Prof. Houda Kawas

https://en.wikipedia.org/wiki/Restriction_fragment_length_polymorphism

https://en.wikipedia.org/wiki/Terminal_restriction_fragment_length_polymorphism

https://biology.stackexchange.com/questions/54292/what-is-the-difference-between-rflp-and-dna-fingerprinting

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1563681/

https://www.quora.com/What-are-the-differences-between-RFLPs-STRs-and-SNPs

https://www.quora.com/What-are-the-main-differences-between-PCR-and-RFLP

https://answers.yahoo.com/question/index?qid=20110624202145AA62WuL

http://www.majordifferences.com/2013/02/difference-between-rapd-and-rflp.html#.WQODKzZWGC4

https://books.google.com/books?id=il9WEQPUfk8C&pg=PA8&lpg=PA8&dq=difference+between+RFLP+and+T-RFLP&source=bl&ots=YiBxWUsrvN&sig=rjMt2UBu73Two2e-ZRxLQi7QhzE&hl=ar&sa=X&ved=0ahUKEwiDw_2y3cfTAhUBCSwKHRVrAdQ4ChDoAQg6MAM#v=onepage&q=difference%20between%20RFLP%20and%20T-RFLP&f=false

https://books.google.com/books?id=uE7_Jja6q4sC&pg=PA428&lpg=PA428&dq=difference+between+RFLP+and+T-RFLP&source=bl&ots=SK2rX-JEKP&sig=w49uSaOgla-SVp-JSeoTbY3AatU&hl=ar&sa=X&ved=0ahUKEwiDw_2y3cfTAhUBCSwKHRVrAdQ4ChDoAQg-MAQ#v=onepage&q=difference%20between%20RFLP%20and%20T-RFLP&f=false

Hoping this will be helpful to your project, good luck

http://www3.appliedbiosystems.com/cms/groups/mcb_marketing/documents/generaldocuments/cms_042272.pdf

https://books.google.com/books?id=dNrTwo5PEUIC&pg=PA94&lpg=PA94&dq=difference+between+RFLP+and+T-RFLP&source=bl&ots=munIi-QhGq&sig=VeZroOP14hVdJa_nIrwx3Oy5g9g&hl=ar&sa=X&ved=0ahUKEwiDw_2y3cfTAhUBCSwKHRVrAdQ4ChDoAQhSMAc#v=onepage&q=difference%20between%20RFLP%20and%20T-RFLP&f=false

http://bmcbioinformatics.biomedcentral.com/articles/10.1186/s12859-014-0360-8

http://www.protocol-online.org/biology-forums/posts/12743.html

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Hi Pooja

You can read the references in attachment 1

https://www.majordifferences.com/2013/02/difference-between-rapd-and-rflp.html

2

https://www.quora.com/What-are-the-differences-between-RFLPs-STRs-and-SNPs

3

https://www.quora.com/What-are-the-main-differences-between-PCR-and-RFLP

4

https://www.soest.hawaii.edu/oceanography/zij/ocn750lectures/ocn750-20051103-dgge

5

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1563681/

6

https://privatewww.essex.ac.uk/~svist/RO_page/t-rflp%2031-3-05.ppt

7

https://books.google.com/books?isbn=0387745203

8

https://bmcbioinformatics.biomedcentral.com/articles/10.1186/s12859-014-0360-8

9

https://docs.appliedbiosystems.com/pebiodocs/00115132.pdf

10

thank you all.

i have DNA sample, isolated from plant root. it may possible, this DNA sample contains various fungal DNA, bacterial DNA including plant DNA and i have to identify only endophytic fungal DNA from DNA sample. so i want to know that  T-RFLP technique is ok ?