# Team:Newcastle/DNA re-hydration

(Difference between revisions)
 Revision as of 12:41, 26 October 2010 (view source)← Older edit Latest revision as of 23:21, 27 October 2010 (view source)Yessa (Talk | contribs) (→Method) (10 intermediate revisions not shown) Line 1: Line 1: {{Team:Newcastle/mainbanner}} {{Team:Newcastle/mainbanner}} - [[Image:Newcastle OSS.JPG|right|200px|Oligonucleotide Specification Sheet (OSS) with the calculations used to work out the volumes of rehydrated primer and H2O required for this specific primer]] + [[Image:Newcastle OSS.JPG|right|200px| Oligonucleotide Specification Sheet (OSS) with the calculations used to work out the volumes of rehydrated primer and H2O required for this specific primer]] =DNA Re-hydration= =DNA Re-hydration= Line 10: Line 10: * Oligonucleotide Specification Sheet (OSS) * Oligonucleotide Specification Sheet (OSS) * OD260 values * OD260 values - * Quantity of DNA in bases + * Sterile Pure Lab Distilled Water - * Pure Lab Distilled Water + - ==Methods== + ==Method== - # To begin, read the Oligonucleotide Specification Sheet to find the OD260 values and Quantity of DNA bases for each primer/DNA. + # To begin, read the Oligonucleotide Specification Sheet to find the OD260 values and number of DNA bases for each primer/DNA. - # Once those values are found, divide the (OD260 value) by (Quantity of DNA bases x10). + # Once those values are found, divide the (OD260 value) by (number of DNA bases x10). # Multiple the result by 1000 # Multiple the result by 1000 - # Divide 10 by the previous result, and multiple all by 100: the end result is the volume of primer/DNA in µl. + # Divide 10 by the previous result, and multiple all by 100: the end result is the volume of primer DNA (in µl) required to prepare 100µl at 10pM concentration. - # Add Pure Lab Distilled Water to make the solution up to 100 µl in total. + # Rehydrate lyopholsed primers in 1ml sterile water - + # Remove the appropiate volume of rehydrated primer and add Pure Lab Distilled Water to make the solution up to 100 µl in total. Please note: Please note: Line 28: Line 27: [[Image:Newcastle Pure Lab Flex 2.JPG|200px]] [[Image:Newcastle Pure Lab Flex 2.JPG|200px]] + + '''Figure 1''' showing Pure Lab Distilled Water apparatus. + + '''Go back to our [[Team:Newcastle/Protocol list|Protocol List]]''' {{Team:Newcastle/footer}} {{Team:Newcastle/footer}}

# DNA Re-hydration

This protocol is used for re-hydration of DNA. We used this mainly for our newly arrived primers.

## Materials required

• Oligonucleotide Specification Sheet (OSS)
• OD260 values
• Sterile Pure Lab Distilled Water

## Method

1. To begin, read the Oligonucleotide Specification Sheet to find the OD260 values and number of DNA bases for each primer/DNA.
2. Once those values are found, divide the (OD260 value) by (number of DNA bases x10).
3. Multiple the result by 1000
4. Divide 10 by the previous result, and multiple all by 100: the end result is the volume of primer DNA (in µl) required to prepare 100µl at 10pM concentration.
5. Rehydrate lyopholsed primers in 1ml sterile water
6. Remove the appropiate volume of rehydrated primer and add Pure Lab Distilled Water to make the solution up to 100 µl in total.