What is the principle behind heating DNA at 94 degree in PCR?
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When a double stranded DNA (dsDNA) molecule is heated to 94oC, the paired strands will separate (denature), by breakage of hydrogen bonding present between complimentary bases, resulting the single stranded DNA formation. This allows the primers access to the single stranded DNA (ssDNA) templates.
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ᴅᴜʀɪɴɢ ᴛʜɪs sᴛᴀɢᴇ ᴛʜᴇ ᴄᴏᴄᴋᴛᴀɪʟ ᴄᴏɴᴛᴀɪɴɪɴɢ ᴛʜᴇ ᴛᴇᴍᴘʟᴀᴛᴇ ᴅɴᴀ ᴀɴᴅ ᴀʟʟ ᴛʜᴇ ᴏᴛʜᴇʀ ᴄᴏʀᴇ ɪɴɢʀᴇᴅɪᴇɴᴛs ɪs ʜᴇᴀᴛᴇᴅ ᴛᴏ 94-95⁰ᴄ. ᴛʜᴇ ʜɪɢʜ ᴛᴇᴍᴘᴇʀᴀᴛᴜʀᴇ ᴄᴀᴜsᴇs ᴛʜᴇ ʜʏᴅʀᴏɢᴇɴ ʙᴏɴᴅs? ʙᴇᴛᴡᴇᴇɴ ᴛʜᴇ ʙᴀsᴇs ɪɴ ᴛᴡᴏ sᴛʀᴀɴᴅs ᴏғ ᴛᴇᴍᴘʟᴀᴛᴇ ᴅɴᴀ ᴛᴏ ʙʀᴇᴀᴋ ᴀɴᴅ ᴛʜᴇ ᴛᴡᴏ sᴛʀᴀɴᴅs ᴛᴏ sᴇᴘᴀʀᴀᴛᴇ.
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