Why is complete digestion necessary in restriction mapping?Restriction mapping में complete digestion क्यों आवश्यक है?
Answer and explanationउत्तर और व्याख्या
Correct answer:सही उत्तर: Because partial digestion can give a false fragment patternBecause partial digestion can give a false fragment pattern
Restriction mapping infers the arrangement of restriction sites from the sizes and number of DNA fragments observed after digestion. The method therefore requires the enzyme to cut every accessible copy of each target site under the selected conditions. If digestion is partial, some molecules are cut at only a subset of sites, producing larger intermediate fragments in addition to the true complete-digest fragments. These extra bands can be mistaken for genuine map products and lead to an incorrect estimate of site order or distance. Thus option A is correct. DNA is not burned, gel electrophoresis is normally essential for observing sizes, and the protein nature of the enzyme does not explain the requirement. Appropriate enzyme amount, incubation, buffer, and DNA quality help obtain complete digestion.Restriction mapping में restriction sites की व्यवस्था का अनुमान digestion के बाद दिखाई देने वाले DNA fragments की संख्या और size से लगाया जाता है। इसलिए चुनी गई conditions में enzyme को प्रत्येक accessible target site को सभी DNA molecules पर काटना चाहिए। यदि digestion partial हो, तो कुछ molecules केवल कुछ sites पर कटेंगे और complete-digest fragments के साथ बड़े intermediate fragments भी बनेंगे। ये extra bands वास्तविक map products समझे जा सकते हैं और site order या distances का गलत अनुमान दे सकते हैं। इसलिए विकल्प A सही है। DNA को जलाना आवश्यक नहीं है, fragment sizes देखने के लिए gel electrophoresis उपयोगी है और enzyme का protein होना इस requirement का कारण नहीं है।
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