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kokoioi
how MMLV RT can end labeling of DNA?It is a RNA-dependent DNA polymerase,but i read some paper use it to label dna which after Restrict enzyme digestion?? expect your reply.thx
Yes :) I think it needs to be double stranded though..Originally posted by kokoioi
hi~Monique
thx for your reply.
It is mean MMLV RT can use DNA for template to synthesis new strand?
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I am not entirely sure of your question. Are you asking whether MMLV RT needs a primer to extend the DNA?Originally posted by kokoioi
and the cleavagesite of XbaI
5'-T^C T A G A-3'
3'-A G A T C^T-5'
(the experiment is for location )
so i think MMLV RT may don't primer when it use DNA for template.
what do you think? and i can't find about MMLV RT use to label DNA in my college.
5'- C T A G A N N N N -3'
3'- T N N N N -5'
MMLV RT stands for Moloney Murine Leukemia Virus Reverse Transcriptase, which is an enzyme commonly used in molecular biology experiments to convert RNA into complementary DNA (cDNA). In DNA end labeling, MMLV RT is used to add a fluorescent or radioactive label to the end of a DNA molecule, allowing for easier detection and visualization.
MMLV RT is highly specific and can accurately label the ends of DNA molecules, making it a reliable method for studying DNA sequences. It is also relatively easy to use and does not require specialized equipment.
Yes, MMLV RT can be used to label both single-stranded and double-stranded DNA. However, it is important to note that the labeling efficiency may vary depending on the type of DNA being labeled.
Yes, there are alternative methods for DNA end labeling such as T4 DNA ligase or terminal deoxynucleotidyl transferase (TdT). These methods may be more suitable for certain applications, so it is important to consider the specific needs of the experiment before choosing a labeling method.
If you are having trouble labeling your DNA using MMLV RT, some potential solutions include optimizing the reaction conditions (such as temperature and incubation time), using higher concentrations of the enzyme, or trying a different labeling method. It may also be helpful to consult with experienced researchers or refer to published protocols for guidance.