8. PCR Table. You need extracted from a different species of rockfish). The tota

Question

8. PCR Table. You need extracted from a different species of rockfish). The total volume of eac to amplify a gene called VIKING from nine different genomic DNA samples (each h PCR is 20 uL, and you will need a master mix to save time variability among. a master mix to save tim e and minimize variability among the reactions. In preparing the master mix, you must include a fudge factor' reaction to compensate for volume loss during pipetting Complete the table below to show the amounts of each component in a single PCR and al master mix. Be sure to list the total volume of your master mix. (5 points) lso in your Component Stock concentration Final concentration or Volume for 1 reaction Volume to add to the master mix amount PCR Buffer 10X MgCl2 Forward 1x 1.5 mM 1 uM 25 mM 10 M Primer Reverse 10 M 1 uM Primer dNTPs Genomic 10 mM 1 mM 11g/mL 50 pg/mL DNA Taq Polymerase 5 U/uL 1 U Nuclease- free Water Total 20

Explanation / Answer

Answer:

Based on the given information, below is the table with details of the amount of the components that should be included to get the desired final concentration and volume.

**Please note:

Genomic DNA sample dilution

Fudge factor consideration: Instead of multiplying the reaction volume of each component for a single reaction with 9 (to account for 9 different reactions), multiply with 9.5 instead to compensate for any loss due to pipetting.

Component Stock Concentration Final Concentration or amount Volume for 1 reaction Volume to add to the master mix (9 reactions) Volume to add to the master mix (with Fudge Factor - 9.5 reactions) PCR Buffer 10X 1X 2 uL 18 uL 19 uL MgCl2 25 mM 1.5 mM 1.2 uL 10.8 uL 11.4 uL Forward Primer 10 uM 1 uM 2 uL 18 uL 19 uL Reverse Primer 10 uM 1 uM 2 uL 18 uL 19 uL dNTPs 10 mM 1 mM 2 uL 18 uL 19 uL Genomic DNA 1 ng/mL (prepared from 1ug/mL) 50 pg/mL 1 uL 9 uL 9.5 uL Taq Polymerase 5 U/uL 1 U 0.2 ul 1.8 uL 1.9 uL Nuclease-free Water 9.6 uL 86.4 uL 91.2 uL Total 20 uL 180 uL 190 uL

Related Questions

Navigate

• Browse (All)
• Browse 8
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.